Cch1 Restores Intracellular Ca2+ in Fungal Cells during Endoplasmic Reticulum Stress
Open accessAbstract
Pathogens endure and proliferate during infection by exquisitely coping with the many stresses imposed by the host to prevent pathogen survival. Recent evidence has shown that fungal pathogens and yeast respond to insults to the endoplasmic reticulum (ER) by initiating Ca(2+) influx across their plasma membrane. Although the high affinity Ca(2+) channel, Cch1, and its subunit Mid1, have been suggested as the protein complex responsible for mediating Ca(2+) influx, a direct demonstration of the gating mechanism of the Cch1 channel remains elusive. In this first mechanistic study of Cch1 channel activity we show that the Cch1 channel from the model human fungal pathogen, Cryptococcus neoformans, is directly activated by the depletion of intracellular Ca(2+) stores. Electrophysiological analysis revealed that agents that enable ER Ca(2+) store depletion promote the development of whole cell inward Ca(2+) currents through Cch1 that are effectively blocked by La(3+) and dependent on the presence of Mid1. Cch1 is permeable to both Ca(2+) and Ba(2+); however, unexpectedly, in contrast to Ca(2+) currents, Ba(2+) currents are steeply voltage-dependent. Cch1 maintains a strong Ca(2+) selectivity even in the presence of high concentrations of monovalent ions. Single channel analysis indicated that Cch1 channel conductance is small, similar to that reported for the Ca(2+) current I(CRAC). This study demonstrates that Cch1 functions as a store-operated Ca(2+)-selective channel that is gated by intracellular Ca(2+) depletion. The inability of cryptococcal cells that lacked the Cch1-Mid1 channel to survive ER stress suggests that Cch1 and its co-regulator, Mid1, are critical players in the restoration of Ca(2+) homeostasis.
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Written from the paper’s full text.
TL;DR
This study provides the first electrophysiological characterization of the *Cryptococcus neoformans* Cch1–Mid1 channel, showing that Cch1 is a store-operated, Ca²⁺-selective channel activated by depletion of intracellular Ca²⁺ stores. The channel is required for cryptococcal survival during ER stress, as mutants lacking Cch1 or Mid1 fail to grow under ER-stressing drugs in low extracellular Ca²⁺.
Key findings
- Co-expression of Cch1 and Mid1 in HEK293 cells produced inward Ca²⁺ currents upon passive ER Ca²⁺ store depletion with 10 µM BAPTA-AM or 100 µM thapsigargin; currents required both subunits and were not seen with Cch1 alone, Mid1 alone, or empty plasmid.
- Cch1 currents were blocked by 10 µM La³⁺, which reduced time-dependent inward Ca²⁺ currents by 75%, but not by physiological concentrations of nifedipine or verapamil.
- Cch1 was permeable to both Ca²⁺ and Ba²⁺, but Ba²⁺ currents were steeply voltage-dependent: at −40 mV, Ca²⁺ current amplitude was ~50% of that at −80 mV, whereas Ba²⁺ current was only ~10% of the −80 mV level. Single-channel Ba²⁺ conductance was ~0.08 pS in 10 mM Ba²⁺.
- Reversal potentials were +42 ± 2 mV for Ba²⁺ inward current in 5 mM Ba²⁺ and +58 ± 2 mV for Ca²⁺ inward current in 5 mM Ca²⁺; Na⁺ conductance was negligible in the absence of divalent cations, indicating strong Ca²⁺ selectivity.
- *C. neoformans* cch1Δ, mid1Δ, and cch1Δmid1Δ mutants were not viable when exposed to tunicamycin, fluconazole, or ketoconazole in low extracellular Ca²⁺ (~500 nM), whereas wild type grew.
Methods
- Heterologous expression: *C. neoformans* CCH1 and MID1 cDNAs were cloned into mammalian expression vectors and transiently transfected into HEK293 cells; expression and plasma membrane localization were assessed by RT-PCR, confocal microscopy, Western blotting, and surface biotinylation.
- Electrophysiology: conventional whole-cell and outside-out single-channel patch clamp recordings measured Cch1 currents activated by BAPTA-AM or thapsigargin, with Ca²⁺ or Ba²⁺ as charge carriers and with monovalent ion substitution; voltage ramps were from −120 to +60 mV.
- Fungal survival: wild type, mid1Δ, cch1Δ, and cch1Δmid1Δ *C. neoformans* strains were serially diluted and spotted on YPD agar with ER-stressing agents (tunicamycin 75 ng/ml, fluconazole 8 µg/ml, ketoconazole 180 ng/ml) with or without cell-impermeable BAPTA.
Limitations
- The paper states that the mechanism by which Mid1 regulates Cch1 function remains unknown, and that Mid1 does not itself conduct Ca²⁺ currents under the tested store-depleted conditions.
- The paper notes that Cch1 could not be detected in surface biotinylation experiments, possibly due to steric hindrance from its large predicted structure, although Mid1 biotinylation and Cch1–Mid1 co-localization supported surface expression.
- The paper states that direct patch clamp of *C. neoformans* spheroplasts was burdened by technical difficulties and inconsistent Cch1 measurements, motivating the heterologous HEK293 system; this means the electrophysiological findings are from a non-fungal expression system.
- Evident from the text: the study examines one fungal species, *C. neoformans*, and does not resolve the structural basis of store-dependent gating or the identity of additional proteins potentially involved in Mid1-dependent regulation.
Model-written summary. Check it against the paper before relying on it.
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Claims the paper makes
0 of 55 cited sources checked · 0 supported · 0 flagged
- Findingown data
Transcripts of Cch1 and Mid1 were not detected in nontransfected HEK293 cells.
“Transcripts of Cch1 and Mid1 were not detected in nontransfected HEK293 cells.”
Quote in the paperSubstantial claim - Findingown data
Agents that enable ER Ca2+ store depletion promote the development of whole cell inward Ca2+ currents through Cch1.
“Electrophysiological analysis revealed that agents that enable ER Ca 2؉ store depletion promote the development of whole cell inward Ca 2؉ currents through Cch1 that are effectively blocked by La 3؉ and dependent on the presence of Mid1.”
Quote in the paperSubstantial claim - Findingown data
Western blot analysis of isolated plasma membranes from HEK293 cells expressing Cch1 or Mid1 revealed polypeptides corresponding to the predicted sizes of Cch1 and Mid1.
“Western blot analysis of isolated plasma membranes from HEK293 cells expressing Cch1 or Mid1 revealed the presence of polypeptides corresponding to the predicted size of Cch1 and Mid1, suggesting that both proteins were indeed sufficiently expressed in HEK cells (Fig. 1C).”
Quote in the paperSubstantial claim - Findingown data
The Cch1 channel from C. neoformans is directly activated by the depletion of intracellular Ca2+ stores.
“In this first mechanistic study of Cch1 channel activity we show that the Cch1 channel from the model human fungal pathogen, Cryptococcus neoformans, is directly activated by the depletion of intracellular Ca 2؉ stores.”
Quote in the paperSubstantial claim - Findingown data
Cch1 channel conductance is small, similar to that reported for the Ca2+ current ICRAC.
“Single channel analysis indicated that Cch1 channel conductance is small, similar to that reported for the Ca 2؉ current I CRAC .”
Quote in the paperNot a substantial claim
- Findingreasoning
Cryptococcal cells that lacked the Cch1-Mid1 channel are unable to survive ER stress.
“The inability of cryptococcal cells that lacked the Cch1-Mid1 channel to survive ER stress suggests that Cch1 and its co-regulator, Mid1, are critical players in the restoration of Ca 2؉ homeostasis.”
- Also stated in: § Abstract · plain
“Consequently, cryptococcal cells that lacked the Cch1 channel could not survive ER stress, suggesting a critical physiological requirement for CMC.”
Quote in the paperSubstantial claim - Also stated in: § Abstract · plain
- Findingown data
Whole cell inward Ca2+ currents through Cch1 are dependent on the presence of Mid1.
“Electrophysiological analysis revealed that agents that enable ER Ca 2؉ store depletion promote the development of whole cell inward Ca 2؉ currents through Cch1 that are effectively blocked by La 3؉ and dependent on the presence of Mid1.”
Quote in the paperSubstantial claim - Findingown data
In contrast to Ca2+ currents, Ba2+ currents through Cch1 are steeply voltage-dependent.
“Cch1 is permeable to both Ca 2؉ and Ba 2؉ ; however, unexpectedly, in contrast to Ca 2؉ currents, Ba 2؉ currents are steeply voltage-dependent.”
Quote in the paperNot a substantial claim - Findingown data
Whole cell inward Ca2+ currents through Cch1 are effectively blocked by La3+.
“Electrophysiological analysis revealed that agents that enable ER Ca 2؉ store depletion promote the development of whole cell inward Ca 2؉ currents through Cch1 that are effectively blocked by La 3؉ and dependent on the presence of Mid1.”
Quote in the paperSubstantial claim - Findingown data
Cch1 maintains a strong Ca2+ selectivity even in the presence of high concentrations of monovalent ions.
“Cch1 maintains a strong Ca 2؉ selectivity even in the presence of high concentrations of monovalent ions.”
Quote in the paperSubstantial claim - Findingown data
Cch1 is permeable to both Ca2+ and Ba2+.
“Cch1 is permeable to both Ca 2؉ and Ba 2؉ ; however, unexpectedly, in contrast to Ca 2؉ currents, Ba 2؉ currents are steeply voltage-dependent.”
Quote in the paperNot a substantial claim - Findingreasoning
Ba2+ currents through Cch1 developed slower than Ca2+ currents.
“Notably, Ba 2ϩ currents developed slower than Ca 2ϩ currents.”
Quote in the paperNot a substantial claim - Findingown data
cch1Δ and mid1Δ single null mutants and cch1Δmid1Δ double mutants were not viable when exposed to tunicamycin in low extracellular [Ca2+].
“The cch1⌬ and mid1⌬ single null mutants and the cch1⌬mid1⌬ double mutants were not viable when exposed to tunicamycin in low extracellular [Ca 2ϩ ] (Fig. 4, B and F).”
Quote in the paperSubstantial claim - Findingown data
Store depletion-mediated inward Ca2+ currents were not detected in HEK cells expressing Cch1 alone, Mid1 alone, or an empty plasmid.
“Store depletion-mediated inward Ca 2ϩ currents were not detected in HEK cells that expressed either Cch1 or Mid1 alone or an empty plasmid, indicating that the activation of these Ca 2ϩ currents was completely dependent on the co-expression of Cch1 and Mid1 and that Cch1 activity required Mid1, consistent with previous genetic evidence (Fig. 2, F-H) (12,14,20,21).”
Quote in the paperSubstantial claim - Findingown data
Ca2+ store depletion resulted in the development of robust, time-dependent Cch1 channel currents with either Ca2+ or Ba2+ as the charge carrier.
“Ca 2ϩ store depletion resulted in the development of robust, time-dependent Cch1 channel currents with either Ca 2ϩ or Ba 2ϩ as the charge carrier.”
Quote in the paperSubstantial claim - Findingown data
These reversal potentials were far less positive than predicted for conditions where extracellular [Ca2+] and [Ba2+] were about 10^5 times higher than inside the patch pipette.
“These values were far less positive than the predicted E Ba and E Ca values for experimental conditions where extracellular [Ca 2ϩ ] and [Ba 2ϩ ] were about 10 5 times higher than the [Ca 2ϩ ] and [Ba 2ϩ ] inside the patch pipette.”
Quote in the paperSubstantial claim - Findingown data
Some inhibition of Cch1 Ca2+ currents was observed in the presence of nifedipine but only at very high concentrations (10 mM).
“Some inhibition of Cch1 Ca 2ϩ currents was observed in the presence of nifedipine but only at very high concentrations (10 mM), suggesting that CnCch1 probably lacks the classic binding site for dihydropyridines and phenylalkyamines.”
Quote in the paperNot a substantial claim - Findingown data
Nifedipine and verapamil did not block Cch1-mediated currents at physiological concentrations.
“The addition of the well characterized Ca 2ϩ channel blockers such as nifedipine and verapamil did not block Cch1-mediated currents at physiological concentrations (data not shown).”
Quote in the paperSubstantial claim - Findingown data
Mid1 did not conduct any Ca2+ currents on its own during passive Ca2+ store depletion.
“Importantly, Mid1 did not conduct any Ca 2ϩ currents on its own during passive Ca 2ϩ store depletion.”
Quote in the paperSubstantial claim - Findingown data
Cch1 could not be detected in Western blots from biotinylated HEK cells expressing Cch1.
“Unfortunately, Cch1 could not be detected in Western blots from biotinylated HEK cells expressing Cch1 (Fig. 1D).”
Quote in the paperNot a substantial claim - Findingown data
The amplitude of Cch1-mediated currents at -40 mV with Ca2+ as the charge carrier was about 50% less than the amplitude at -80 mV.
“The amplitude of Cch1-mediated currents at Ϫ40 mV with Ca 2ϩ as the charge carrier was ϳ50% less than the amplitude measured at Ϫ80 mV (Fig. 3B).”
Quote in the paperNot a substantial claim - Findingown data
Depletion of Ca2+ stores by BAPTA-AM produced Cch1-mediated currents with either Ca2+ or Ba2+ as the charge carrier.
“Macroscopic I-V curves demonstrated that depletion of Ca 2ϩ stores by BAPTA-AM produced Cch1-mediated currents with either Ca 2ϩ or Ba 2ϩ as the charge carrier.”
Quote in the paperSubstantial claim - Findingown data
Cch1 was highly permeable to barium ions, similar to other Ca2+ channels in higher eukaryotes.
“Similar to other Ca 2ϩ channels in higher eukaryotes, Cch1 was also highly permeable to barium ions.”
Quote in the paperNot a substantial claim - Findingcitation
A similar growth phenotype was observed in the presence of fluconazole or ketoconazole.
“A similar phenotype was observed in the presence of fluconazole or ketoconazole, drugs belonging to the azole class of antifungals (Fig. 4, C, D, G, and H) (30,31).”
Quote in the paperNot a substantial claim0 of 2 sources checked - Findingown data
Addition of 10 μM La3+ resulted in a 75% reduction in the time-dependent inward Ca2+ currents.
“In contrast, the addition of 10 M La 3ϩ resulted in a 75% reduction in the time-dependent inward Ca 2ϩ currents, indicating that La 3ϩ blocked Cch1 channel activity (Fig. 2D).”
Quote in the paperSubstantial claim - Findingown data
Thapsigargin induces time-dependent inward Ca2+ currents through Cch1.
“Consistent with the stimulation of Ca 2ϩ currents by chelation of Ca 2ϩ from the ER, time-dependent inward Ca 2ϩ currents through Cch1 also developed in response to thapsigargin (Fig. 2E).”
Quote in the paperSubstantial claim - Findingown data
With Ba2+, the current amplitude at -40 mV was only about 10% of that at -80 mV, indicating Ba2+ permeation through Cch1 is voltage-dependent.
“However, with Ba 2ϩ the current amplitude at Ϫ40 mV was only ϳ10% of the current levels detected at Ϫ80 mV, indicating that Ba 2ϩ permeation through Cch1 was voltage-dependent.”
Quote in the paperNot a substantial claim - Findingown data
The C-terminal GFP tag on Cch1 did not affect Cch1 Ca2+ or Ba2+ channel currents.
“The Ca 2ϩ or Ba 2ϩ channel currents of Cch1 were not affected by the presence of the C-terminal GFP tag on the Cch1 protein as indicated by the I-V plot (Fig. 3E).”
Quote in the paperNot a substantial claim - Premisecitation
Strains of C. neoformans lacking CCH1 or MID1 were not viable in conditions of limiting extracellular Ca2+.
“Genetic analysis has indicated that strains of C. neoformans lacking CCH1 or MID1 were not viable in conditions of limiting extracellular [Ca 2ϩ ], consistent with its role as the only high affinity Ca 2ϩ channel in the plasma membrane (7,14).”
Quote in the paperSubstantial claim0 of 2 sources checked - Interpretationreasoning
Cch1 functions primarily to restore Ca2+ homeostasis.
“suggesting that Cch1 is gated by the passive depletion of intracellular Ca 2ϩ and that it functions primarily to restore Ca 2ϩ homeostasis.”
Quote in the paperSubstantial claim - Premisecitation
C. neoformans survival in low extracellular Ca2+ environments depends on the combined activity of Cch1 and Mid1.
“Its ability to survive in low [Ca 2ϩ ] environments is dependent on the combined activity of Cch1, the predicted pore of the channel complex, and Mid1, a binding partner of Cch1 (7,14,20).”
Quote in the paperSubstantial claim0 of 3 sources checked - Premisecitation
Treatment of fungal cells with azole antifungals led to an influx of extracellular Ca2+ that promoted the survival of fungi exposed to azole-induced stress.
“Treatment of fungal cells with azole antifungals led to an influx of extracellular Ca 2ϩ that promoted the survival of fungi exposed to azole-induced stress (16 -19).”
Quote in the paperSubstantial claim0 of 4 sources checked- cites [16] Kaur, 2004, Antimicrob. Agents Chemother. source
Not checked in the free run.
- cites [17] Bonilla, 2003, Mol. Biol. Cell. source
Not checked in the free run.
- cites [18] Bonilla, 2002, EMBO source
Not checked in the free run.
- cites [19] Edlind, 2002, Mol. Microbiol. source
Not checked in the free run.
- Interpretationreasoning
Cch1 and Mid1 are critical players in the restoration of Ca2+ homeostasis.
“The inability of cryptococcal cells that lacked the Cch1-Mid1 channel to survive ER stress suggests that Cch1 and its co-regulator, Mid1, are critical players in the restoration of Ca 2؉ homeostasis.”
Quote in the paperSubstantial claim - Study design and conductown data
Detection of mRNA transcripts of Cch1 and Mid1 confirmed the expression of the channel in HEK293 cells that were previously transfected with CCH1 and MID1.
“Detection of mRNA transcripts of Cch1 and Mid1 confirmed the expression of the channel in HEK293 cells that were previously transfected with CCH1 and MID1 (Fig. 1A).”
Quote in the paperSubstantial claim - Premisecitation
Cch1 is the only high affinity Ca2+ channel in the plasma membrane of C. neoformans.
“Genetic analysis has indicated that strains of C. neoformans lacking CCH1 or MID1 were not viable in conditions of limiting extracellular [Ca 2ϩ ], consistent with its role as the only high affinity Ca 2ϩ channel in the plasma membrane (7,14).”
Quote in the paperSubstantial claim0 of 2 sources checked - Premisecitation
Measurement of Cch1 channel activity in a heterologous expression system had been impossible until recently because of the significant challenge posed by molecular cloning of CCH1 due to toxicity of the CCH1 gene or gene product in E. coli cells.
“Unfortunately, the measurement of Cch1 channel activity in a heterologous expression system had been impossible until recently because of the significant challenge posed by the molecular cloning of CCH1 as a result of the toxicity of the CCH1 gene or gene product in Escherichia coli cells (8).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [8] Paidhungat, 1997, Mol. Cell. Biol. source
Not checked in the free run.
- Interpretationcitation
Mid1 likely forms an oligomer through sulfide bonding similar to Mid1 in yeast.
“This result suggested that Mid1 likely forms an oligomer through sulfide bonding similar to Mid1 in yeast (40).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [40] Yoshimura, 2004, Exp. Cell Res. source
Not checked in the free run.
- Interpretationown data
La3+ blocks Cch1 channel activity.
“In contrast, the addition of 10 M La 3ϩ resulted in a 75% reduction in the time-dependent inward Ca 2ϩ currents, indicating that La 3ϩ blocked Cch1 channel activity (Fig. 2D).”
Quote in the paperSubstantial claim - Interpretationreasoning
The La3+-induced block of Ca2+ currents further strengthens the functional similarity between CMC and SOC channels in mammalian cells.
“further strengthening the functional similarity between CMC and SOC channels in mammalian cells.”
Quote in the paperSubstantial claim - Interpretationown data
CnCch1 probably lacks the classic binding site for dihydropyridines and phenylalkyamines.
“suggesting that CnCch1 probably lacks the classic binding site for dihydropyridines and phenylalkyamines.”
Quote in the paperNot a substantial claim - Premisecitation
At about 500 nM free Ca2+, low affinity Ca2+ channels and/or transporters allow Ca2+ influx that supports viable CMC mutants.
“Lowering [Ca 2ϩ ] further rendered the CMC mutants inviable, but at ϳ500 nM, low affinity Ca 2ϩ channels and/or transporters allow for Ca 2ϩ influx that supports viable CMC mutants (7,20).”
Quote in the paperSubstantial claim0 of 2 sources checked - Study design and conductown data
Biotinylation of Mid1 confirms its expression on the surface of HEK293 cells.
“Nevertheless, the biotinylation of Mid1 confirms its expression on the surface of HEK293 cells, and the co-localization of Mid1 with Cch1 strongly suggests that Cch1 is also expressed on the surface of HEK cells.”
Quote in the paperSubstantial claim - Interpretationcitation
The voltage dependence of Ba2+ influx is probably due to very hyperpolarized potentials needed to displace Ba2+ bound to the Cch1 pore, likely due to weak electrostatic repulsion between Ba2+ ions.
“This suggested that very hyperpolarized potentials were probably needed to displace Ba 2ϩ bound to the Cch1 pore with an incoming Ba 2ϩ ion likely due to weak electrostatic repulsion between Ba 2ϩ ions, thus resulting in the voltage dependence of Ba 2ϩ influx (25).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [25] Hille, 1992
Not checked in the free run.
- Interpretationown data
Cch1 activity plays an essential role in survival of cryptococcal cells in response to ER stress by coupling depleted ER Ca2+ stores to Ca2+ influx across the plasma membrane.
“Here, we show that Cch1 activity plays an essential role in the survival of cryptococcal cells in response to ER stress by coupling the depleted state of ER Ca 2ϩ stores to the influx of Ca 2ϩ across the plasma membrane.”
Quote in the paperSubstantial claim - Premisecitation
The La3+-induced block of Ca2+ currents is a pharmacological hallmark of SOC channels.
“The La 3ϩ -induced block of Ca 2ϩ currents is a pharmacological hallmark of SOC channels (3, 4), further strengthening the functional similarity between CMC and SOC channels in mammalian cells.”
Quote in the paperSubstantial claim0 of 2 sources checked - Interpretationcitation
The slower development of Ba2+ currents is probably due to secondary effects of Ba2+ blocking K+ conductances and affecting the electrical potential difference across the plasma membrane, thus reducing the driving force for Ba2+ influx.
“This was probably due to secondary effects of Ba 2ϩ blocking K ϩ conductances and affecting the electrical potential difference across the plasma membrane thus reducing the driving force for Ba 2ϩ influx (Fig. 2, A and B) (25).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [25] Hille, 1992
Not checked in the free run.
- Interpretationreasoning
Co-localization of Mid1 with Cch1 strongly suggests that Cch1 is also expressed on the surface of HEK cells.
“the co-localization of Mid1 with Cch1 strongly suggests that Cch1 is also expressed on the surface of HEK cells.”
Quote in the paperSubstantial claim - Interpretationreasoning
The SOC gating mechanism of Cch1 indicates that Cch1 functions primarily to replenish depleted Ca2+ levels and reestablish Ca2+ homeostasis.
“In summary, the SOC gating mechanism of Cch1 indicates that the Cch1 channel functions primarily to replenish depleted Ca 2ϩ levels and reestablish Ca 2ϩ homeostasis.”
Quote in the paperSubstantial claim - Premisecitation
Previous genetic data demonstrated identical phenotypic defects of strains lacking either Mid1 or Cch1, including nonviability in low [Ca2+] medium.
“This is consistent with previous genetic data that demonstrated identical phenotypic defects of strains lacking either Mid1 or Cch1 including nonviability in low [Ca 2ϩ ] medium (7,11,15,20).”
Quote in the paperNot a substantial claim0 of 3 sources checked
- May combine two claims. 32
- Classified as probably not a substantial claim. 17
May combine two claims.
FindingcitationMid1 is expressed predominantly on the cell surface of HEK293 cells and co-localizes with Cch1.
“Cells expressing a Mid1-GFP fusion protein revealed a cell surface distribution of Mid1 that co-localized with Cch1 (Fig. 1B) (24).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [24] Vergara, 1998, Curr. Opin. Neurobiol. source
Not checked in the free run.
May combine two claims.
FindingcitationUnder nonreducing conditions, two distinct Mid1 protein bands were detected: a 100 kDa monomer and a 200 kDa likely complex.
“The 100 kDa band represented the Mid1 monomer, and the 200 kDa band likely corresponded to a Mid1 complex (Fig. 1C) (40).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [40] Yoshimura, 2004, Exp. Cell Res. source
Not checked in the free run.
May combine two claims.
Findingown dataBa2+ currents through Cch1 were negligible at membrane potentials depolarized beyond -60 mV, whereas Ca2+ currents were detected at these potentials, reflecting voltage independence of Ca2+ currents under physiological conditions.
“We found that Ba 2ϩ currents were negligible at membrane potentials depolarized beyond Ϫ60 mV, whereas Ca 2ϩ currents were detected at these same potentials, reflecting the voltage independence of Ca 2ϩ currents through Cch1 under physiological conditions.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataSingle channel Ba2+ currents through Cch1 are voltage-dependent: no events at -60 mV, increased activity at -80 mV.
“Single channel events were not detected at Ϫ60 mV with Ba 2ϩ as the charge carrier; however, channel activity increased dramatically at Ϫ80 mV, indicating the voltage dependence of single channel Ba 2ϩ currents through Cch1 (Fig. 3C).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataInward Ca2+ currents through Cch1 developed promptly in response to BAPTA-AM and spontaneously declined to resting levels.
“Inward Ca 2ϩ currents through Cch1 developed promptly in response to BAPTA-AM and spontaneously declined to resting levels (Fig. 2C).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataAs membrane potentials became decreasingly hyperpolarized, Ba2+ currents fell more than Ca2+ currents, resulting in more pronounced inward rectification.
“As membrane potentials became decreasingly hyperpolarized, Ba 2ϩ currents fell to a greater extent than Ca 2ϩ currents, resulting in a more pronounced inward rectification (Fig. 3A).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataSingle Cch1 channels exhibited long duration openings with a conductance of about 0.08 picosiemens in 10 mM Ba2+.
“Single channels exhibited long duration openings with a conductance of ϳ0.08 picosiemens in 10 mM Ba 2ϩ , consistent with the small channel conductance reported for CRAC channels (27).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [27] Kerschbaum, 1999, Science source
Not checked in the free run.
May combine two claims.
Findingown dataNa+ conductance through Cch1 in the absence of divalent cations was negligible, and addition of Ca2+ in the presence of Na+ resulted in large inward currents with a shift in Erev to positive potentials approaching ECa.
“Interestingly, Na ϩ conductance through Cch1 in the absence of divalent cations was negligible, and the addition of Ca 2ϩ in the presence of Na ϩ resulted in large inward currents with a shift in E rev to positive potentials approaching E Ca (Fig. 3D).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataCurrents were not detected in HEK cells expressing empty plasmids, suggesting the Ca2+ currents detected were mediated by Cch1.
“In addition, currents were not detected in HEK cells expressing empty plasmids, strongly suggesting that the Ca 2ϩ currents detected were mediated by Cch1 (Fig. 3E).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataCch1 was more selective for Ca2+ than Ba2+ because Erev was more positive and closer to the equilibrium potential for Ca2+.
“However, Cch1 was more selective for Ca 2ϩ than Ba 2ϩ because E rev was more positive and closer to the equilibrium potential for Ca 2ϩ .”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataThe reversal potential of the Ba2+ inward current in 5 mM Ba2+ was +42 ± 2 mV, and for the Ca2+ inward current in 5 mM Ca2+ was +58 ± 2 mV.
“The reversal potential of the Ba 2ϩ inward current (E Ba ) in 5 mM Ba 2ϩ was ϩ42 Ϯ 2 mV and ϩ58 Ϯ 2 mV for the Ca 2ϩ inward current (E Ca ) in 5 mM Ca 2ϩ (Fig. 3A).”
Quote in the paperSubstantial claimMay combine two claims.
FindingreasoningBa2+ movement through Cch1 was voltage-dependent, whereas Ca2+ influx was not.
“Unexpectedly, Ba 2ϩ movement through Cch1 was voltage-dependent; however, Ca 2ϩ influx was not.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataIn the presence of tunicamycin, wild type C. neoformans showed no significant growth defect except the mid1Δ mutant; strains lacking a functional CMC had severely compromised growth.
“In the presence of the ER-stressing agent tunicamycin, a wild type strain of C. neoformans showed no significant growth defect with the exception of the mid1⌬ mutant (Fig. 4, A and E); however, the growth of C. neoformans strains lacking a functional CMC was severely compromised.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataAlthough Ca2+ and Ba2+ were both permeable through Cch1, Ba2+ currents were steeply voltage-dependent.
“Surprisingly, although Ca 2ϩ and Ba 2ϩ were both permeable, Ba 2ϩ currents were steeply voltage-dependent.”
Quote in the paperNot a substantial claimMay combine two claims.
FindingcitationUnder Ca2+-depleted conditions, Mid1 did not promote influx of divalent cations when expressed alone, indicating Mid1 does not have independent channel activity under these conditions.
“Under Ca 2ϩ -depleted conditions, Mid1 did not promote the influx of divalent cations when expressed on its own, indicating that Mid1 does not have any independent channel activity under these particular conditions (39).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [39] Kanzaki, 1999, Science source
Not checked in the free run.
May combine two claims.
PremisecitationStresses imposed on the ER can promote a prolonged depletion of Ca2+ that will lead to cell death unless ER Ca2+ levels are replenished.
“Stresses imposed on the ER can promote a prolonged depletion of Ca 2ϩ that will lead to cell death unless ER Ca 2ϩ levels are replenished (1)(2)(3).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites (1)(2)(3)
Not checked in the free run.
Classified as probably not a substantial claim.
PremisecitationStore-operated Ca2+ (SOC) entry is a process whereby Ca2+ influx across the eukaryotic plasma membrane results from the depletion of Ca2+ from endoplasmic reticulum (ER) stores.
“Store-operated Ca 2ϩ (SOC) 2 entry is a process whereby Ca 2ϩ influx across the eukaryotic plasma membrane results from the depletion of Ca 2ϩ from endoplasmic reticulum (ER) stores (1)(2)(3)(4)(5)(6).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites (1)(2)(3)(4)(5)(6)
Not checked in the free run.
Classified as probably not a substantial claim.
PremisecitationChanges in secretory Ca2+ levels promoted the influx of extracellular 45Ca2+ possibly through CMC in yeast.
“This notion was reinforced by experiments demonstrating that changes in secretory Ca 2ϩ levels promoted the influx of extracellular 45 Ca 2ϩ possibly through CMC in yeast (20);”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [20] Locke, 2000, Mol. Cell. Biol. source
Not checked in the free run.
May combine two claims.
Interpretationown dataCch1 functions as a store-operated Ca2+-selective channel that is gated by intracellular Ca2+ depletion.
“This study demonstrates that Cch1 functions as a store-operated Ca 2؉ -selective channel that is gated by intracellular Ca 2؉ depletion.”
- Also stated in: § Abstract · hedged
“suggesting that Cch1 is gated by the passive depletion of intracellular Ca 2ϩ and that it functions primarily to restore Ca 2ϩ homeostasis.”
Quote in the paperSubstantial claim- Also stated in: § Abstract · hedged
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationAll lower eukaryotes including C. neoformans express the Cch1-Mid1 channel (CMC) in their plasma membrane.
“All lower eukaryotes including the model fungal pathogen, Cryptococcus neoformans, express the Cch1-Mid1 channel (CMC) in their plasma membrane (7)(8)(9)(10)(11).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites (7)(8)(9)(10)(11)
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationC. neoformans is a primary human pathogen because it causes life-threatening meningitis primarily in immunocompromised patients.
“C. neoformans is a primary human pathogen because it causes life-threatening meningitis primarily in immunocompromised patients (12,13).”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites [12] Mitchell, 1995, Clin. Microbiol. Rev. source
Not checked in the free run.
- cites [13] Casadevall, 1998
Not checked in the free run.
Classified as probably not a substantial claim.
PremisecitationResolving channel gating is best accomplished by patch clamp techniques because these techniques can determine specific kinetic parameters of ion channels such as channel selectivity, conductance, and activation kinetics.
“Resolving channel gating is best accomplished by patch clamp techniques because these techniques can successfully determine specific kinetic parameters of ion channels such as channel selectivity, channel conductance, and activation kinetics (21).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [21] Hamill, 1981, Pflugers Arch. Eur. J. Physiol. source
Not checked in the free run.
May combine two claims.
PremisecitationER stress can be detrimental to fungal cells if ER function is not restored because the ER is key in protein processing and Ca2+ signaling.
“ER stress can be detrimental to fungal cells if ER function is not restored because in addition to its crucial role in the posttranslational processing of proteins, the ER is also a key player in Ca 2ϩ signaling (32).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [32] Brostrom, 2003, Cell Calcium source
Not checked in the free run.
May combine two claims.
Interpretationown dataCryptococcal cells required CMC to survive ER stress conditions, supporting that Cch1 plays an essential role in reestablishing Ca2+ homeostasis.
“These results indicated that cryptococcal cells required CMC to survive ER stress conditions and supported the notion that the Cch1 channel plays an essential role in reestablishing Ca 2ϩ homeostasis.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationIn C. neoformans, mediators of Ca2+ signaling such as calcineurin, calmodulin, and SERCA are central to maintaining intracellular Ca2+ homeostasis while withstanding host stresses.
“In C. neoformans mediators of the Ca 2ϩ signaling pathway, such as calcineurin, calmodulin, and the sarco (endo)plasmic reticulum Ca 2ϩ -ATPase are central to maintaining intracellular Ca 2ϩ homeostasis while withstanding the multitude of stresses imposed by the host (33).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [33] Fan, 2007, Infect. Immun. source
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenThe Cch1 channel protein was tagged with a C-terminal GFP instead of an N-terminal GFP to avoid interfering with the localization of Cch1 to the plasma membrane of HEK293 cells.
“The Cch1 channel protein was tagged with a C-terminal GFP instead of an N-terminal GFP to avoid interfering with the localization of Cch1 to the plasma membrane of HEK293 cells.”
Quote in the paperNot a substantial claimMay combine two claims.
Interpretationown dataThe Cch1-Mid1 proteins constituted a functional Ca2+ channel in HEK293 cells that exhibited some features inherent to non-voltage-gated SOC channels.
“The results presented here demonstrate that the Cch1-Mid1 proteins constituted a functional Ca 2ϩ channel in HEK293 cells that exhibited some features inherent to the non-voltage-gated SOC channels.”
Quote in the paperSubstantial claim- Rests on an assumption: The heterologous HEK293 cell system faithfully reports the functional properties of the Cch1-Mid1 channel as it operates in C. neoformans.
May combine two claims.
Interpretationown dataCch1 functions primarily to replenish ER Ca2+ stores and reestablish Ca2+ homeostasis.
“The presence of a specific Ca 2ϩ chelator and an inhibitor of the Ca 2ϩ -ATPase, both capable of depleting intracellular ER Ca 2ϩ stores, led to the development of Ca 2ϩ inward currents at negative membrane potentials, indicating that Cch1 functions primarily to replenish these stores and reestablish Ca 2ϩ homeostasis.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown dataThe cch1⌬mid1⌬ mutant strain was confirmed by PCR and Southern blot analysis.
“The cch1⌬mid1⌬ mutant strain was confirmed by PCR and Southern blot analysis.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenAll voltages have been corrected for liquid junction potentials.
“All voltages have been corrected for liquid junction potentials.”
Quote in the paperNot a substantial claimMay combine two claims.
Interpretationown dataCch1-mediated store depletion-activated Ca2+ currents require co-expression of Cch1 and Mid1, i.e., Cch1 activity requires Mid1.
“Store depletion-mediated inward Ca 2ϩ currents were not detected in HEK cells that expressed either Cch1 or Mid1 alone or an empty plasmid, indicating that the activation of these Ca 2ϩ currents was completely dependent on the co-expression of Cch1 and Mid1 and that Cch1 activity required Mid1, consistent with previous genetic evidence (Fig. 2, F-H) (12,14,20,21).”
Quote in the paperSubstantial claim0 of 3 sources checkedProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductnone givenFollowing blue/white colony selection, positive transformants were isolated and sequenced.
“Following blue/white colony selection, positive transformants were isolated and sequenced.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationSOC channels are highly permeable to sodium and other monovalent ions in the absence of divalent cations.
“One of the distinguishing properties of SOC channels is their highly permeability to sodium ions and other monovalent ions in the absence of divalent cations (27).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [27] Kerschbaum, 1999, Science source
Not checked in the free run.
May combine two claims.
InterpretationcitationUnlike some SOC channels in higher eukaryotes that are highly permeable to Na+ in the absence of divalent cations, Cch1 is weakly permeable to Na+ and maintains strong Ca2+ selectivity even in high [Na+].
“These results indicated that unlike some SOC channels in higher eukaryotes that are highly permeable to Na ϩ in the absence of divalent cations, Cch1 is weakly permeable to Na ϩ and maintains a strong Ca 2ϩ selectivity even in the presence of high [Na ϩ ] (4, 27).”
Quote in the paperSubstantial claim0 of 2 sources checkedMay combine two claims.
Interpretationown dataCs+ carried some outward current through Cch1 and significantly contributed to the reversal potential by shifting EBa and ECa to less positive values.
“These results indicated that Cs ϩ carried some outward current through Cch1 and in doing so made a significant contribution to the reversal potential of Cch1 by shifting the E Ba and E Ca to less positive values.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationCRAC channels also exhibit a similar voltage-dependent influx of Ba2+ at negative membrane potentials.
“Interestingly, CRAC channels, a subtype of the voltage-independent SOC channels in mammalian cells, also exhibit a similar voltage-dependent influx of Ba 2ϩ at negative membrane potentials (34).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [34] Bakowski, 2007, Cell Calcium source
Not checked in the free run.
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenAll data were corrected for leak currents.
“All data were corrected for leak currents.”
Quote in the paperNot a substantial claimMay combine two claims.
InterpretationcitationElectrophysiological evidence supports a Ca2+ depletion gating mechanism for Cch1 and a role for Cch1 in SOC entry in lower eukaryotes.
“The electrophysiological evidence provided here strongly supports a Ca 2ϩ depletion gating mechanism for Cch1 and further substantiates a role for Cch1 in the process of SOC entry in lower eukaryotes (20).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [20] Locke, 2000, Mol. Cell. Biol. source
Not checked in the free run.
Classified as probably not a substantial claim.
PremisecitationIn eukaryotes, the ER constitutes a finite reservoir of Ca2+ primarily mobilized for signaling that must be replenished to sustain signaling and restore Ca2+ homeostasis.
“It is known that in eukaryotes the ER constitutes a finite reservoir of Ca 2ϩ primarily mobilized for signaling that must be replenished to sustain the duration of signaling and ultimately restore Ca 2ϩ homeostasis in the ER and the cytosol (1-3, 32).”
Quote in the paperSubstantial claim0 of 4 sources checkedMay combine two claims.
Interpretationown dataCch1 is a Ca2+-selective SOC channel gated by depletion of intracellular Ca2+.
“This first mechanistic study of Cch1 indicates that it is a Ca 2ϩ -selective SOC channel that is gated by the depletion of intracellular Ca 2ϩ based on the following observations: (i) inward Ca 2ϩ currents were activated upon the passive depletion of Ca 2ϩ with BAPTA-AM and thapsigargin at negative membrane potentials; (ii) Cch1 was permeable to Ba 2ϩ ions; and (iii) Ca 2ϩ currents were blocked by”
Quote in the paperSubstantial claim- Rests on an assumption: The heterologous HEK293 cell system faithfully reports the functional properties of the Cch1-Mid1 channel as it operates in C. neoformans.
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenA nontagged version of a Cch1 plasmid was constructed and used as a control to assess the effect of the GFP fusion on the kinetics of Cch1.
“A nontagged version of a Cch1 plasmid was also constructed and used as a control to assess the effect of the GFP fusion on the kinetics of Cch1.”
Quote in the paperNot a substantial claimMay combine two claims.
PremisecitationThapsigargin depletes ER Ca2+ stores by inhibiting the ER Ca2+-ATPase pump, preventing reuptake as Ca2+ passively leaks from the ER.
“Thapsigargin, an inhibitor of the ER Ca 2ϩ -ATPase pump, leads to the depletion of Ca 2ϩ stores because as Ca 2ϩ passively leaks from the ER, Ca 2ϩ levels are no longer balanced by reuptake (26).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [26] Rogers, 1995, Biosci. Rep. source
Not checked in the free run.
May combine two claims.
Interpretationown dataActivation of Cch1-mediated Ca2+ currents required the presence of Mid1, indicating Cch1 cannot promote Ca2+ influx independently of Mid1.
“The activation of Cch1-mediated Ca 2ϩ currents required the presence of Mid1, indicating that Cch1 cannot promote Ca 2ϩ influx independently of Mid1.”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationown dataMid1 is not involved in trafficking of Cch1 to the plasma membrane because Cch1 was expressed in the plasma membrane of HEK293 cells independently of Mid1.
“Our results rule out the possibility that Mid1 is involved in the trafficking of Cch1 to the plasma membrane because Cch1 was clearly expressed in the plasma membrane of HEK293 cells independently of Mid1.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S4 (based).
InterpretationreasoningThis enables C. neoformans to overcome ER stress-mediated events that are likely rampant during infection and colonization of the host.
“This enables C. neoformans to overcome ER stress-mediated events that are likely rampant during infection and colonization of the host.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationThe NALCN neuronal Na+ channel, which like CnCch1 lacks a bona fide voltage sensor, was found to be voltage-independent.
“Moreover, the recently characterized NALCN neuronal Na ϩ channel, which like CnCch1 has fewer positively charged amino acids (lysine and arginine) within its voltage-sensing S4 segment and thus lacks a bona fide voltage sensor, was found to be voltage-independent (38).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [38] Lu, 2007, Cell source
Not checked in the free run.
May combine two claims.
PremisecitationRecent evidence demonstrated that localization of Cch1 to the plasma membrane in cryptococcal cells is dependent on elongation factor EF3, suggesting Mid1 is excluded from this process.
“In addition, recent evidence has demonstrated that the localization of Cch1 to the plasma membrane in cryptococcal cells is dependent on an elongation factor (EF3), further suggesting that Mid1 is excluded from this process (14).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [14] Liu, 2008, Eukaryot. Cell source
Not checked in the free run.
May combine two claims.
InterpretationcitationCch1 is not voltage-gated, supported by previous findings that Cch1 in C. neoformans lacks the conserved voltage sensor motif.
“Our results indicate that Cch1 is not voltage-gated, and this is further supported by our previous findings demonstrating that the protein sequence of Cch1 in C. neoformans lacks the highly conserved voltage sensor motif, the hallmark of all voltage-gated channels in eukaryotic cells (7,(35)(36)(37).”
Quote in the paperSubstantial claim0 of 3 sources checked
The paper relies on this without support.
The heterologous HEK293 expression system faithfully recapitulates the functional properties of the native Cch1-Mid1 channel in C. neoformans.
“Consequently, the expression of Cch1 in a heterologous expression system such as the HEK293 (human embryonic kidney) mammalian cell line appeared to be the best approach that would permit direct functional analysis of the Cch1 channel.”
“To resolve Ca 2ϩ currents through CMC by patch clamp techniques, HEK293 cells were co-transfected with CCH1 and MID1 (21).”
The paper relies on this without support.
The inward Ca2+ currents measured in transfected HEK293 cells are mediated by Cch1-Mid1 rather than endogenous channels.
“To measure currents through Cch1, HEK293 cells expressing Cch1 and Mid1 were visually selected based on bright GFP fluorescence.”
“Ca 2ϩ store depletion resulted in the development of robust, time-dependent Cch1 channel currents with either Ca 2ϩ or Ba 2ϩ as the charge carrier.”
The paper relies on this without support.
The heterologous HEK293 cell system faithfully reports the functional properties of the Cch1-Mid1 channel as it operates in C. neoformans.
“The results presented here demonstrate that the Cch1-Mid1 proteins constituted a functional Ca 2ϩ channel in HEK293 cells that exhibited some features inherent to the non-voltage-gated SOC channels.”
The paper relies on this without support.
The growth-inhibitory effects of tunicamycin, fluconazole, and ketoconazole in the spot assays are due to ER stress.
“In the presence of the ER-stressing agent tunicamycin, a wild type strain of C. neoformans showed no significant growth defect with the exception of the mid1⌬ mutant (Fig. 4, A and E); however, the growth of C. neoformans strains lacking a functional CMC was severely compromised.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Under ER stress, Ca2+ in the ER may leak into the cytoplasm, potentially increasing Ca2+ availability within mitochondria.
“Under ER stress, Ca2+ in the ER may leak into the cytoplasm, potentially resulting in an increase in the availability of Ca2+ within the mitochondria35,36.”
From Impairing the interaction between Erg11 and cytochrome P450 reductase Ncp1 enhances azoles’ antifungal activities (2025) · cites this paper as [36]
The section “Cch1 Is Activated by the Depletion of Intracellular Ca²⁺” (page not provided) says Ca²⁺ passively leaks from the ER when thapsigargin prevents its reuptake. The paper does not say that this leakage increases Ca²⁺ availability in mitochondria, so that part of the attributed claim is unsupported. (LLM, conf=0.87)
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Lanthanum charge differs between results and figure caption
MinorThe results identify the channel blocker as La 3ϩ, whereas the caption for the same experiment identifies it as La 2ϩ.
“the addition of 10 M La 3ϩ resulted in a 75% reduction in the time-dependent inward Ca 2ϩ currents”
“D, 10 M La 2ϩ in the bath solution significantly reducing Ca 2ϩ currents through Cch1.”
Cites
Hamill, O. P., Marty, A., Neher, E., Sakmann, B., & Sigworth, F. J. (1981) · Pflügers Archiv - European Journal of Physiology · doi:10.1007/bf00656997 · Cited by 18,436 · see citing works
Ionic channels of excitable membranes
(1985) · General Pharmacology The Vascular System · doi:10.1016/0306-3623(85)90172-7 · Cited by 7,133 · see citing works
Ionic Channels of Excitable Membranes
Keynes, R. D. (1985) · Trends in Neurosciences · doi:10.1016/0166-2236(85)90088-8 · Cited by 2,600 · see citing works
A model for receptor-regulated calcium entry
Putney, J. W. (1986) · Cell Calcium · doi:10.1016/0143-4160(86)90026-6 · Cited by 2,575 · see citing works
Structure and Regulation of Voltage-Gated Ca <sup>2+</sup> Channels
Catterall, W. A. (2000) · Annual Review of Cell and Developmental Biology · doi:10.1146/annurev.cellbio.16.1.521 · Cited by 2,344 · see citing works
A mutation in Orai1 causes immune deficiency by abrogating CRAC channel function
Feske, S., Gwack, Y., Prakriya, M., Srikanth, S., Puppel, S.-H., Tanasă, B., Hogan, P. G., Lewis, R. S., Daly, M. J., & Rao, A. (2006) · Nature · doi:10.1038/nature04702 · Cited by 2,278 · see citing works
Tsien, R. Y. (1980) · Biochemistry · doi:10.1021/bi00552a018 · Cited by 2,243 · see citing works
Store-Operated Calcium Channels
Parekh, A. B., & Putney, J. W. (2005) · Physiological Reviews · doi:10.1152/physrev.00057.2003 · Cited by 2,089 · see citing works
Capacitative calcium entry revisited
Putney, J. W. (1990) · Cell Calcium · doi:10.1016/0143-4160(90)90016-n · Cited by 1,487 · see citing works
Zhang, S. L., Yu, Y., Roos, J., Kozak, J. A., Deerinck, T. J., Ellisman, M. H., Stauderman, K. A., & Cahalan, M. D. (2005) · Nature · doi:10.1038/nature04147 · Cited by 1,374 · see citing works
Cryptococcosis in the era of AIDS--100 years after the discovery of Cryptococcus neoformans
Mitchell, T. G., & Perfect, J. R. (1995) · Clinical Microbiology Reviews · doi:10.1128/cmr.8.4.515 · Cited by 1,213 · see citing works
Antifungal agents: mechanisms of action
Odds, F. C., Brown, A. J. P., & Gow, N. A. R. (2003) · Trends in Microbiology · doi:10.1016/s0966-842x(03)00117-3 · Cited by 1,202 · see citing works
Voltage Sensor of Kv1.2: Structural Basis of Electromechanical Coupling
Long, S. B., Campbell, E. B., & MacKinnon, R. (2005) · Science · doi:10.1126/science.1116270 · Cited by 982 · see citing works
Calcium-activated potassium channels
Vergara, C., Latorre, R., Marrion, N. V., & Adelman, J. P. (1998) · Current Opinion in Neurobiology · doi:10.1016/s0959-4388(98)80056-1 · Cited by 954 · see citing works
Lu, B., Su, Y., Das, S., Liu, J., Xia, J., & Ren, D. (2007) · Cell · doi:10.1016/j.cell.2007.02.041 · Cited by 381 · see citing works
Essential role of calcineurin in response to endoplasmic reticulum stress
Bonilla, M., Nastase, K. K., & Cunningham, K. W. (2002) · The EMBO Journal · doi:10.1093/emboj/21.10.2343 · Cited by 255 · see citing works
Warnock, D. W. (1999) · Journal of Antimicrobial Chemotherapy · doi:10.1093/jac/44.1.139 · Cited by 232 · see citing works
Iida, H., Nakamura, H., Ono, T., Okumura, M. S., & Anraku, Y. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.12.8259 · Cited by 218 · see citing works
Molecular Identification of a Eukaryotic, Stretch-Activated Nonselective Cation Channel
Kanzaki, M., Nagasawa, M., Kojima, I., Sato, C., Naruse, K., Sokabe, M., & Iida, H. (1999) · Science · doi:10.1126/science.285.5429.882 · Cited by 213 · see citing works
Locke, E. G., Bonilla, M., Liang, L., Takita, Y., & Cunningham, K. W. (2000) · Molecular and Cellular Biology · doi:10.1128/mcb.20.18.6686-6694.2000 · Cited by 212 · see citing works
Brostrom, M. A., & Brostrom, C. O. (2003) · Cell Calcium · doi:10.1016/s0143-4160(03)00127-1 · Cited by 197 · see citing works
Gene Disruption by Biolistic Transformation in Serotype D Strains of Cryptococcus neoformans
Davidson, R. C., Cruz, M. C., Sia, R. A. L., Allen, B., Alspaugh, J. A., & Heitman, J. (2000) · Fungal Genetics and Biology · doi:10.1006/fgbi.1999.1180 · Cited by 194 · see citing works
Paidhungat, M., & Garrett, S. (1997) · Molecular and Cellular Biology · doi:10.1128/mcb.17.11.6339 · Cited by 191 · see citing works
The <i>Saccharomyces cerevisiae</i> CCH1 gene is involved in calcium influx and mating
Fischer, M., Schnell, N., Chattaway, J., Davies, P., Dixon, G., & Sanders, D. (1997) · FEBS Letters · doi:10.1016/s0014-5793(97)01466-x · Cited by 185 · see citing works
Kaur, R., Castaño, I., & Cormack, B. P. (2004) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.48.5.1600-1613.2004 · Cited by 167 · see citing works
Bonilla, M., & Cunningham, K. W. (2003) · Molecular Biology of the Cell · doi:10.1091/mbc.e03-02-0113 · Cited by 165 · see citing works
Use of thapsigargin to study Ca2+ homeostasis in cardiac cells
Rogers, T. B., Inesi, G., Wade, R., & Lederer, W. J. (1995) · Bioscience Reports · doi:10.1007/bf01788366 · Cited by 147 · see citing works
Single-Channel Recording of a Store-Operated Ca <sup>2+</sup> Channel in Jurkat T Lymphocytes
Kerschbaum, H. H., & Cahalan, M. D. (1999) · Science · doi:10.1126/science.283.5403.836 · Cited by 134 · see citing works
Zelter, A., Benčina, M., Bowman, B. J., Yarden, O., & Read, N. D. (2004) · Fungal Genetics and Biology · doi:10.1016/j.fgb.2004.05.001 · Cited by 129 · see citing works
Dissecting ICRAC, a store-operated calcium current
Hogan, P. G., & Rao, A. (2007) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2007.03.009 · Cited by 117 · see citing works
Antifungal activity in <i>Saccharomyces cerevisiae</i> is modulated by calcium signalling
Edlind, T., Smith, L., Henry, K., Katiyar, S., & Nickels, J. (2002) · Molecular Microbiology · doi:10.1046/j.1365-2958.2002.03165.x · Cited by 114 · see citing works
Marks, P. W., & Maxfield, F. R. (1991) · Analytical Biochemistry · doi:10.1016/0003-2697(91)90044-t · Cited by 102 · see citing works
Liu, M., Du, P., Heinrich, G., Cox, G. M., & Gelli, A. (2006) · Eukaryotic Cell · doi:10.1128/ec.00158-06 · Cited by 76 · see citing works
Self-citationHallen‐Adams, H. E., & Trail, F. (2007) · Eukaryotic Cell · doi:10.1128/ec.00248-07 · Cited by 71 · see citing works
Clinical efficacy of new antifungal agents
Kauffman, C. A. (2006) · Current Opinion in Microbiology · doi:10.1016/j.mib.2006.08.001 · Cited by 64 · see citing works
Fan, W., Idnurm, A., Breger, J., Mylonakis, E., & Heitman, J. (2007) · Infection and Immunity · doi:10.1128/iai.01977-06 · Cited by 57 · see citing works
Models of the Structure and Voltage-Gating Mechanism of the Shaker K+ Channel
Durell, S. R., Shrivastava, I. H., & Guy, H. R. (2004) · Biophysical Journal · doi:10.1529/biophysj.104.040618 · Cited by 56 · see citing works
Yoshimura, H., Tada, T., & Iida, H. (2004) · Experimental Cell Research · doi:10.1016/j.yexcr.2003.09.020 · Cited by 45 · see citing works
Bakowski, D., & Parekh, A. B. (2007) · Cell Calcium · doi:10.1016/j.ceca.2007.01.005 · Cited by 23 · see citing works
Vu, K., Bautos, J. M., Hong, M.-P., & Gelli, A. (2009) · Analytical Biochemistry · doi:10.1016/j.ab.2009.06.039 · Cited by 15 · see citing works
Self-citationLiu, M., & Gelli, A. (2008) · Eukaryotic Cell · doi:10.1128/ec.00116-08 · Cited by 15 · see citing works
Self-citation
Flagged first, then by citations
Research works printed but not linked: 1
- Iida, 1994, Mol. Cell. Biol., 14, 8259
Cited by
Sorted by citations
Regulation of Cation Balance inSaccharomyces cerevisiae
Cyert, M., & Philpott, C. C. (2013) · Genetics · doi:10.1534/genetics.112.147207 · Cited by 253 · see citing works
Sodium Leak Channels in Neuronal Excitability and Rhythmic Behaviors
Ren, D. (2011) · Neuron · doi:10.1016/j.neuron.2011.12.007 · Cited by 167 · see citing works
New Regulators of a High Affinity Ca2+ Influx System Revealed through a Genome-wide Screen in Yeast
Martin, D. C., Kim, H., Mackin, N. A., Maldonado‐Báez, L., Evangelista, C. C., Beaudry, V. G., Dudgeon, D. D., Naiman, D. Q., Erdman, S. E., & Cunningham, K. W. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.177451 · Cited by 93 · see citing works
Lange, M., & Peiter, E. (2020) · Frontiers in Microbiology · doi:10.3389/fmicb.2019.03100 · Cited by 90 · see citing works
Orm Proteins Integrate Multiple Signals to Maintain Sphingolipid Homeostasis
Gururaj, C. S., Federman, R. S., & Chang, A. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.472860 · Cited by 72 · see citing works
Cation Transporters of Candida albicans—New Targets to Fight Candidiasis?
Volkova, M. B., Atamas, A., Tsarenko, A., Rogachev, A. V., & Guskov, A. (2021) · Biomolecules · doi:10.3390/biom11040584 · Cited by 68 · see citing works
Wang, S., Cao, J., Liu, X., Hu, H., Shi, J. R., Zhang, S., Keller, N. P., & Lü, L. (2012) · PLoS ONE · doi:10.1371/journal.pone.0046564 · Cited by 64 · see citing works
Liebeskind, B. J., Hillis, D. M., & Zakon, H. H. (2012) · Molecular Biology and Evolution · doi:10.1093/molbev/mss182 · Cited by 64 · see citing works
The Involvement of the Mid1/Cch1/Yvc1 Calcium Channels in Aspergillus fumigatus Virulence
de Castro, P. A., Chiaratto, J., Winkelströter, L. K., Bom, V. L. P., Ramalho, L. N. Z., Goldman, M. H. S., Brown, N. A., & Goldman, G. H. (2014) · PLoS ONE · doi:10.1371/journal.pone.0103957 · Cited by 60 · see citing works
Ngamskulrungroj, P., Chang, Y., Hansen, B., Bugge, C., Fischer, E., & Kwon-Chung, K. J. (2012) · PLoS ONE · doi:10.1371/journal.pone.0033022 · Cited by 56 · see citing works
Repurposing drugs to fast-track therapeutic agents for the treatment of cryptococcosis
Truong, M., Monahan, L. G., Carter, D. A., & Charles, I. G. (2018) · PeerJ · doi:10.7717/peerj.4761 · Cited by 54 · see citing works
Cavinder, B., & Trail, F. (2012) · Eukaryotic Cell · doi:10.1128/ec.00007-12 · Cited by 48 · see citing works
Tisi, R., Rigamonti, M. M., Groppi, S., & Belotti, F. (2016) · AIMS molecular science · doi:10.3934/molsci.2016.4.505 · Cited by 40 · see citing works
Zhang, Y., Zheng, Q., Sun, C., Song, J., Gao, L., Zhang, S., Muñoz, A., Read, N. D., & Lü, L. (2016) · PLoS Genetics · doi:10.1371/journal.pgen.1005977 · Cited by 39 · see citing works
Identification and Analysis of Cation Channel Homologues in Human Pathogenic Fungi
Prole, D. L., & Taylor, C. W. (2012) · PLoS ONE · doi:10.1371/journal.pone.0042404 · Cited by 37 · see citing works
Pozdnyakov, I. A., Matantseva, O., & Skarlato, S. O. (2018) · Scientific Reports · doi:10.1038/s41598-018-21897-7 · Cited by 36 · see citing works
Stefan, C. P., Zhang, N., Sokabe, T., Rivetta, A., Slayman, C. L., Montell, C., & Cunningham, K. W. (2013) · Eukaryotic Cell · doi:10.1128/ec.00299-12 · Cited by 32 · see citing works
Cai, X. (2012) · The Journal of Membrane Biology · doi:10.1007/s00232-012-9415-9 · Cited by 32 · see citing works
Robust Internal Elastic Lamina Fenestration in Skeletal Muscle Arteries
Kirby, B. S., Bruhl, A., Sullivan, M., Francis, M., Dinenno, F. A., & Earley, S. (2013) · PLoS ONE · doi:10.1371/journal.pone.0054849 · Cited by 30 · see citing works
Ding, X., Yu, Q., Xu, N., Wang, Y., Cheng, X., Qian, K., Zhao, Q., Zhang, B., Xing, L., & Li, M. (2013) · Fungal Genetics and Biology · doi:10.1016/j.fgb.2013.05.010 · Cited by 30 · see citing works
Troppens, D. M., Chu, M., Holcombe, L. J., Gleeson, O., O’Gara, F., Read, N. D., & Morrissey, J. P. (2013) · Fungal Genetics and Biology · doi:10.1016/j.fgb.2013.04.006 · Cited by 28 · see citing works
Zhang, S., Zheng, H., Chen, Q., Chen, Y., Wang, S., Lü, L., & Zhang, S. (2017) · Applied and Environmental Microbiology · doi:10.1128/aem.00673-17 · Cited by 26 · see citing works
Ca2+ homeostasis in the budding yeast Saccharomyces cerevisiae: Impact of ER/Golgi Ca2+ storage
D’hooge, P., Coun, C., Eyck, V. V., Faes, L., Ghillebert, R., Mariën, L., Winderickx, J., & Callewaert, G. (2015) · Cell Calcium · doi:10.1016/j.ceca.2015.05.004 · Cited by 25 · see citing works
Zhang, D., Ma, Z., Kai, K., Hu, T., Bi, W., Yang, Y., Shi, W. M., Wang, Z., & Ye, Y. (2023) · Scientia Horticulturae · doi:10.1016/j.scienta.2023.112091 · Cited by 24 · see citing works
Cch1p Mediates Ca2+ Influx to Protect Saccharomyces cerevisiae against Eugenol Toxicity
Roberts, S. K., McAinsh, M. R., & Widdicks, L. (2012) · PLoS ONE · doi:10.1371/journal.pone.0043989 · Cited by 22 · see citing works
Hong, M.-P., Vu, K., Bautos, J. M., Tham, R. C., Jamklang, M., Uhrig, J. P., & Gelli, A. (2012) · Eukaryotic Cell · doi:10.1128/ec.00130-12 · Cited by 22 · see citing works
Self-citationXu, D., Cheng, J., Cao, C., Wang, L., & Jiang, L. (2015) · FEMS Yeast Research · doi:10.1093/femsyr/fov079 · Cited by 18 · see citing works
Revel, B., Catty, P., Ravanel, S., Bourguignon, J., & Alban, C. (2021) · Journal of Hazardous Materials · doi:10.1016/j.jhazmat.2021.126894 · Cited by 17 · see citing works
The virally encoded killer proteins from Ustilago maydis
Allen, A., Islamovic, E., Kaur, J., Gold, S. E., Shah, D. M., & Smith, T. J. (2012) · Fungal Biology Reviews · doi:10.1016/j.fbr.2012.10.001 · Cited by 16 · see citing works
Vu, K., Bautos, J. M., & Gelli, A. (2015) · Eukaryotic Cell · doi:10.1128/ec.00100-15 · Cited by 16 · see citing works
Self-citationStefan, C. P., & Cunningham, K. W. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.508705 · Cited by 15 · see citing works
Gonçalves, A. P., Cordeiro, M., Monteiro, J., Muñoz, A., Correia‐de‐Sá, P., Read, N. D., & Videira, A. (2014) · Journal of Cell Science · doi:10.1242/jcs.152058 · Cited by 15 · see citing works
Valproate inhibits MAP kinase signalling and cell cycle progression in S. cerevisiae
Desfossés-Baron, K., Hammond-Martel, I., Simoneau, A., Sellam, A., Roberts, S., & Würtele, H. (2016) · Scientific Reports · doi:10.1038/srep36013 · Cited by 11 · see citing works
Iida, K., Teng, J., Cho, T., Yoshikawa-Kimura, S., & Iida, H. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.m117.810283 · Cited by 11 · see citing works
Li, W., Whiteway, M., Hang, S., Yu, J., Lu, H., & Jiang, Y. (2025) · Nature Communications · doi:10.1038/s41467-025-62131-z · Cited by 13 · see citing works
“Under ER stress, Ca 2+ in the ER may leak into the cytoplasm, potentially resulting in an increase in the availability of Ca 2+ within the mitochondria 35,36 .”
Chandel, A., & Bachhawat, A. K. (2017) · Journal of Cell Science · doi:10.1242/jcs.202853 · Cited by 10 · see citing works
Ca2+ Signalling Differentially Regulates Germ-Tube Formation and Cell Fusion in Fusarium oxysporum
Kurian, S. M., Lichius, A., & Read, N. D. (2022) · Journal of Fungi · doi:10.3390/jof8010090 · Cited by 9 · see citing works
PpMID1 Plays a Role in the Asexual Development and Virulence of Phytophthora parasitica
Hwu, F.-Y., Lai, M.-W., & Liou, R. (2017) · Frontiers in Microbiology · doi:10.3389/fmicb.2017.00610 · Cited by 9 · see citing works
Qian, H., Chen, Q., Zhang, S., & Lü, L. (2018) · Frontiers in Microbiology · doi:10.3389/fmicb.2018.00977 · Cited by 9 · see citing works
Calcium Ion Channels in Saccharomyces cerevisiae
Dong, X. (2023) · Journal of Fungi · doi:10.3390/jof9050524 · Cited by 7 · see citing works
Coupling of a voltage‐gated Ca2+ channel homologue with a plasma membrane H+‐ATPase in yeast
Cho, T., Aya, I.-K., Fukata, Y., Nakayama, Y., Iida, K., Fukata, M., & Iida, H. (2016) · Genes to Cells · doi:10.1111/gtc.12458 · Cited by 7 · see citing works
Xie, M., Zhou, X., Xia, Y., & Cao, Y. (2019) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-018-09589-8 · Cited by 7 · see citing works
Oyama, M., Tamaki, H., Yamaguchi, Y., Ogita, A., Tanaka, T., & Fujita, K. (2020) · FEMS Yeast Research · doi:10.1093/femsyr/foaa003 · Cited by 7 · see citing works
Lukumay, N. R., Li, Y., Wang, H., & Wei, W. (2025) · Virulence · doi:10.1080/21505594.2025.2569624 · Cited by 5 · see citing works
Wang, Z., Shen, Z.-F., Wang, J.-Y., Cai, Y., Li, L., Liao, J., Lu, J., Zhu, X., Lin, F., & Liu, X. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24119297 · Cited by 5 · see citing works
Hayashi, T., Oishi, K., Kimura, M., Iida, K., & Iida, H. (2020) · Journal of Biological Chemistry · doi:10.1074/jbc.ra120.014378 · Cited by 5 · see citing works
NALCN/Cch1 channelosome subunits originated in early eukaryotes
Senatore, A., Mayorova, T. D., Yáñez-Guerra, L. A., Elkhatib, W., Bejoy, B., Lory, P., & Monteil, A. (2025) · The Journal of General Physiology · doi:10.1085/jgp.202413636 · Cited by 2 · see citing works
Zanni, E., Franco, M., Nakano, M., Iida, H., Palleschi, C., & Uccelletti, D. (2012) · Microbiology · doi:10.1099/mic.0.056283-0 · Cited by 2 · see citing works
Molecular Drug Targets in Candida glabrata
Gupta, P., Rai, N., & Kumar, N. (2017) · Journal of Graphic Era University · Cited by 1 · see citing works
Role of Intracellular Ca2+in Plants and Fungi
Campbell, A. K. (2014) · doi:10.1002/9781118675410.ch9 · Cited by 1 · see citing works
Mulyar, O. A., Teng, A. C. T., & Gramolini, A. O. (2012) · Expert Review of Proteomics · doi:10.1586/epr.11.76
(2014) · Wiley eBooks · doi:10.1002/9781118675410.biblio
Simpson-Lavy, K. J., & Kupiec, M. (2025) · International Journal of Molecular Sciences · doi:10.3390/ijms26052198
Bouasse, M. (2017) · doi:10.70675/6ade06edz19aez4babzb28bzc209b78e30e4
- Authors
- Hong, M.-P., Vu, K., Bautos, J., & Gelli, A.
- Affiliations
- University of California, Davis (US)
- Corresponding author
- Gelli, Angie
- ORCID
- Gelli, Angie
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2010-04
- Volume
- 285
- Issue
- 14
- Pages
- 10951-10958
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC2856300
- OpenAlex
- W2020411557
- Semantic Scholar
- 917cd00888600f02b7589cbb1179a12f7b701923
- Cited by
- 54
- References
- 41
- Citations by year
- 2025: 4 · 2023: 3 · 2022: 1 · 2021: 2 · 2020: 3 · 2019: 1 · 2018: 3 · 2017: 6 · 2016: 4 · 2015: 3 · 2014: 4 · 2013: 6 · 2012: 12
- FWCI
- 3.62
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases (R01AI054477, AI054477, R01AI078848, AI078848)
- Topics
- Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
- Keywords
- MID1, store-operated calcium entry, endoplasmic reticulum stress, Cryptococcus neoformans, intracellular calcium homeostasis, calcium influx, barium currents, single-channel conductance, calcium store depletion
- MeSH terms
- Barium, Calcium, Cell Membrane, Cells, Cultured, Cryptococcosis, Cryptococcus neoformans, Endoplasmic Reticulum, Humans, Kidney, Calcium Channels, Ion Channel Gating, Patch-Clamp Techniques, Biotinylation
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-09.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-09.
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