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PreprintCited by 18Escherichia coli research studiesBacterial Genetics and BiotechnologyPlant Pathogenic Bacteria Studies

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PopB-PcrV Interactions are Essential for Pore Formation in the Pseudomonas aeruginosa Type III Secretion System Translocon

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Kundracik, Emma Caitlin, Trichka, Josephine, Díaz Aponte, José Enrique, Rietsch, Arne·2022·mBio
DOI 10.1101/2022.01.21.477233PMC PMC9600203

Preprint — published as 10.1128/mbio.02381-22 · Paper page

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Abstract

The type III secretion system (T3SS) is a syringe-like virulence factor which delivers bacterial proteins directly into the cytoplasm of host cells. An essential component of the system is the translocon, which creates a pore in the host cell membrane through which proteins are injected. In Pseudomonas aeruginosa , the translocation pore is formed by proteins PopB and PopD and attaches to the T3SS needle via the needle tip protein PcrV. The pore is multimeric, but the exact stoichiometry and structure of the pore are unknown. We took a genetic approach to map contact points within the system by taking advantage of the fact that the translocator proteins of Pseudomonas aeruginosa and the related Aeromonas hydrophila T3SS are incompatible and cannot be freely exchanged. We created chimeric versions of P. aeruginosa PopB and A. hydrophila AopB to intentionally disrupt and restore protein-protein interactions. We identified a chimeric B-translocator that specifically breaks an interaction with the needle tip protein and interferes with the formation of the translocation pore. Breaking the interaction did not disrupt membrane insertion, arguing that the needle tip protein chaperones formation of the translocation pore.

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FIG 1 Translocator proteins from P. aeruginosa and A. hydrophila are not interchangeable. (A) Model of component parts of the translocon (left) and assembled translocation pore (right). PopB is inserted in the membrane with N- and C-termini toward the extracellular space (54). PopD has its C terminus in the extracellular space (25). The pentameric arrangement of PcrV is based on alignment to the Salmonella needle tip structure (55). (B) Amino acid sequence identity and similarity for P. aeruginosa (accession number AE004091) and A. hydrophila AH-3 (accession number AY528667) translocators. Sequences were aligned with ClustalW. (C) AlphaFold2 models of the needle tip protein from P. aeruginosa (PcrV), A. hydrophila strain AH-3 (AcrV), and two chimeric-fusion V proteins. The hybrid Acollar protein has an AcrV substitution in the N-terminal globular domain [residues 1 to F143 from AcrV are fused to PcrV(Q124–end)]. The hybrid Atip contains the bulkier tip domain of AcrV substituted for the tip domain of PcrV (residues 1 to A158 from PcrV, A179 to S310 from AcrV, and V244 to the end from PcrV). Monomer models were created from protein sequences using AlphaFold2 implemented in Google Colab Notebooks (33, 56) and visualized with UCSF ChimeraX (57). The position of the collar domain is uncertain and varies across models. The tip domain has a high degree of confidence and is static between models (55). Pseudomonas segments are shown in purple, and Aeromonas segments in yellow. (D) All strains lack the pore-forming translocator operon (ΔpcrH-popBD) and produce the indicated needle tip protein: PcrV (strain RP3624), AcrV (strain RP7360), Acollar (strain RP6166), or Atip (strain RP6425). These background strains were complemented with plasmids carrying the indicated translocator proteins along with both export chaperone homologs, pcrH and acrH (Table S1). A549 epithelial cells were infected, and cellular rounding (indicating delivery of ExoS) was monitored by microscopy after 2 h. In our analysis, we consider a translocator system nonfunctional if it leads to less than 25% cell rounding at 2 h (dotted line). Statistical significance was calculated by two-way analysis of variance (ANOVA) with the Tukey multiple-comparison test. n = 3 biological replicates. Error bars show standard deviations. ****, P < 0.0001; ns, not statistically significant (P > 0.05). Statistical data for all comparisons are listed in Table S3. Schematic figures were created using BioRender.com.
FIG 1 Translocator proteins from P. aeruginosa and A. hydrophila are not interchangeable. (A) Model of component parts of the translocon (left) and assembled translocation pore (right). PopB is inserted in the membrane with N- and C-termini toward the extracellular space (54). PopD has its C terminus in the extracellular space (25). The pentameric arrangement of PcrV is based on alignment to the Salmonella needle tip structure (55). (B) Amino acid sequence identity and similarity for P. aeruginosa (accession number AE004091) and A. hydrophila AH-3 (accession number AY528667) translocators. Sequences were aligned with ClustalW. (C) AlphaFold2 models of the needle tip protein from P. aeruginosa (PcrV), A. hydrophila strain AH-3 (AcrV), and two chimeric-fusion V proteins. The hybrid Acollar protein has an AcrV substitution in the N-terminal globular domain [residues 1 to F143 from AcrV are fused to PcrV(Q124–end)]. The hybrid Atip contains the bulkier tip domain of AcrV substituted for the tip domain of PcrV (residues 1 to A158 from PcrV, A179 to S310 from AcrV, and V244 to the end from PcrV). Monomer models were created from protein sequences using AlphaFold2 implemented in Google Colab Notebooks (33, 56) and visualized with UCSF ChimeraX (57). The position of the collar domain is uncertain and varies across models. The tip domain has a high degree of confidence and is static between models (55). Pseudomonas segments are shown in purple, and Aeromonas segments in yellow. (D) All strains lack the pore-forming translocator operon (ΔpcrH-popBD) and produce the indicated needle tip protein: PcrV (strain RP3624), AcrV (strain RP7360), Acollar (strain RP6166), or Atip (strain RP6425). These background strains were complemented with plasmids carrying the indicated translocator proteins along with both export chaperone homologs, pcrH and acrH (Table S1). A549 epithelial cells were infected, and cellular rounding (indicating delivery of ExoS) was monitored by microscopy after 2 h. In our analysis, we consider a translocator system nonfunctional if it leads to less than 25% cell rounding at 2 h (dotted line). Statistical significance was calculated by two-way analysis of variance (ANOVA) with the Tukey multiple-comparison test. n = 3 biological replicates. Error bars show standard deviations. ****, P < 0.0001; ns, not statistically significant (P > 0.05). Statistical data for all comparisons are listed in Table S3. Schematic figures were created using BioRender.com.CC BY · PubMed Central
FIG 2 The AopB-Atip interaction is mapped to two small regions of the B-translocator. (A) Schematic diagram of PopB-AopB chimeras. Residues are numbered according to PopB amino acid sequence. PopB transmembrane domains are predicted to be residues 170 to 201 (TM1) and residues 232 to 258 (TM2) (58). Green lines indicate PopB. Yellow bubbles indicate AopB. Transmembrane domains are indicated by white bubbles. (B) Background strains PAO1F ΔexsE ΔexoT exoY ΔpcrH-popBD with wild-type pcrV (strain RP3624) or Atip (strain RP6425) were complemented with acrH, pcrH, and popD and the indicated popB-aopB chimera. A549 cells were infected for 2 h, and the proportion of round cells assessed. n = 3 biological replicates. Error bars show standard deviations. Statistical significance was calculated using two-way ANOVA with the Sidak multiple-comparison test. Significance thresholds: *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.0001; ns, not statistically significant (P > 0.05). (C) Schematic diagram of Bmix chimera with wild-type PcrV or chimeric Atip, illustrating the tip-dependent phenotype. The Bmix/PcrV combination has reduced cytotoxicity, presumably due to incompatibilities introduced by the homologous substitutions (mismatch phenotype). The Bmix/Atip combination performs significantly better in the cytotoxicity assay (rescue phenotype). (D) Cysteine mutations were introduced into PopB and PcrV in the locations shown schematically. In subsequent experiments, PcrV proteins having a single cysteine mutation (Q87C or S202C) were paired with PopB mutants having a single cysteine mutation (I117C or S321C). A variant of PopB includes a 5.3-kDa size tag consisting of four copies of the vesicular stomatitis virus glycoprotein (4×VSV-G) inserted after residue 225 (25). (E) Background strains PAO1F ΔexsE exoS(G/A−) ΔexoT exoY ΔpcrH-popBD with wild-type pcrV (strain RP3670), pcrV(Q87C) (strain RP12661), or pcrV(S202C) (strain RP12598) were complemented with acrH, pcrH, and popD and the indicated version of popB. A549 cells were infected in the presence of copper for 2 h. After infection, off-target disulfide bond formation was blocked by the addition of iodoacetamide. Membrane extract samples were prepared in SDS sample buffer lacking or containing dithiothreitol (DTT) as indicated. The blot is representative of three independent experiments. Molecular-weight size markers (in kilodaltons) are indicated. “exoS(G/A−)” denotes the mutant exoS(R146K/E379D/E381D), which is enzymatically inactive (30). Schematic figures were created using BioRender.com.
FIG 2 The AopB-Atip interaction is mapped to two small regions of the B-translocator. (A) Schematic diagram of PopB-AopB chimeras. Residues are numbered according to PopB amino acid sequence. PopB transmembrane domains are predicted to be residues 170 to 201 (TM1) and residues 232 to 258 (TM2) (58). Green lines indicate PopB. Yellow bubbles indicate AopB. Transmembrane domains are indicated by white bubbles. (B) Background strains PAO1F ΔexsE ΔexoT exoY ΔpcrH-popBD with wild-type pcrV (strain RP3624) or Atip (strain RP6425) were complemented with acrH, pcrH, and popD and the indicated popB-aopB chimera. A549 cells were infected for 2 h, and the proportion of round cells assessed. n = 3 biological replicates. Error bars show standard deviations. Statistical significance was calculated using two-way ANOVA with the Sidak multiple-comparison test. Significance thresholds: *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.0001; ns, not statistically significant (P > 0.05). (C) Schematic diagram of Bmix chimera with wild-type PcrV or chimeric Atip, illustrating the tip-dependent phenotype. The Bmix/PcrV combination has reduced cytotoxicity, presumably due to incompatibilities introduced by the homologous substitutions (mismatch phenotype). The Bmix/Atip combination performs significantly better in the cytotoxicity assay (rescue phenotype). (D) Cysteine mutations were introduced into PopB and PcrV in the locations shown schematically. In subsequent experiments, PcrV proteins having a single cysteine mutation (Q87C or S202C) were paired with PopB mutants having a single cysteine mutation (I117C or S321C). A variant of PopB includes a 5.3-kDa size tag consisting of four copies of the vesicular stomatitis virus glycoprotein (4×VSV-G) inserted after residue 225 (25). (E) Background strains PAO1F ΔexsE exoS(G/A−) ΔexoT exoY ΔpcrH-popBD with wild-type pcrV (strain RP3670), pcrV(Q87C) (strain RP12661), or pcrV(S202C) (strain RP12598) were complemented with acrH, pcrH, and popD and the indicated version of popB. A549 cells were infected in the presence of copper for 2 h. After infection, off-target disulfide bond formation was blocked by the addition of iodoacetamide. Membrane extract samples were prepared in SDS sample buffer lacking or containing dithiothreitol (DTT) as indicated. The blot is representative of three independent experiments. Molecular-weight size markers (in kilodaltons) are indicated. “exoS(G/A−)” denotes the mutant exoS(R146K/E379D/E381D), which is enzymatically inactive (30). Schematic figures were created using BioRender.com.CC BY · PubMed Central
FIG 3 The Bmix-PcrV mismatch does not hinder membrane insertion of PopB. A549 cells were infected for 2 h with P. aeruginosa cells producing PopD and the indicated needle tip/B-translocator combination (background strains RP3670 and RP11222). The control strain lacks pcrH, popB, and popD. After infection, cells were washed with 1 M KCl to remove nonspecifically adhered protein, and membrane-inserted translocator proteins were extracted using Triton X-100 (see Materials and Methods). (A) Translocator proteins, as well as E-cadherin (fractionation/loading control), were detected by Western blotting in membrane extract and supernatant samples. The blot is representative of five independent experiments. The positions of molecular-weight size markers (in kilodaltons) are indicated. (B) Quantification of 5 biological replicates. The amount of PopB/Bmix is normalized to that of E-cadherin or EGFR. Error bars show standard deviations. Statistical significance was assessed with two-way ANOVA using the Sidak multiple-comparison test. ns, not statistically significant (i.e., P > 0.05).
FIG 3 The Bmix-PcrV mismatch does not hinder membrane insertion of PopB. A549 cells were infected for 2 h with P. aeruginosa cells producing PopD and the indicated needle tip/B-translocator combination (background strains RP3670 and RP11222). The control strain lacks pcrH, popB, and popD. After infection, cells were washed with 1 M KCl to remove nonspecifically adhered protein, and membrane-inserted translocator proteins were extracted using Triton X-100 (see Materials and Methods). (A) Translocator proteins, as well as E-cadherin (fractionation/loading control), were detected by Western blotting in membrane extract and supernatant samples. The blot is representative of five independent experiments. The positions of molecular-weight size markers (in kilodaltons) are indicated. (B) Quantification of 5 biological replicates. The amount of PopB/Bmix is normalized to that of E-cadherin or EGFR. Error bars show standard deviations. Statistical significance was assessed with two-way ANOVA using the Sidak multiple-comparison test. ns, not statistically significant (i.e., P > 0.05).CC BY · PubMed Central
FIG 4 Pore formation is disrupted by mismatch between PopB and PcrV. (A, B) A549 cells were infected in the presence of propidium iodide with RP11946 (PAO1 ΔexsE ΔexoSTY pcrV+ ΔpcrH-popBD) or RP11948 [PAO1 ΔexsE ΔexoSTY pcrV::acrV(tip) ΔpcrH-popBD] producing PopD and the indicated translocators. RP2317 (PAO1 ΔexsE ΔexoSTY pcrV+) was the wild-type control. After 2 h, cells were washed and imaged (bright field and fluorescence with Texas red filter). (A) Representative micrographs from one biological replicate are shown. (B) Four biological replicates are quantified. Fluorescent cells were scored manually, and the results of each experiment were normalized to the results for the PopB/PcrV strain. Error bars show standard deviations. Statistical differences were analyzed with two-way ANOVA and the Sidak multiple-comparison test: *, P < 0.05;ns, not statistically significant (P > 0.05). (C) Sheep erythrocytes were pretreated with papain, which cleaves surface glycoproteins and exposes cryptic binding sites for P. aeruginosa hemagglutinin (53). The erythrocytes were infected for 1 h with strain RP3670 [PAO1 ΔexsE exoS(G/A−) pcrV+ ΔpcrH-popBD] or strain RP11222 [PAO1 ΔexsE exoS(G/A−) pcrV::Atip ΔpcrH-popBD] producing PopD and the indicated translocators. The erythrocyte suspension was then mixed, unbroken erythrocytes pelleted, and supernatant UV-Vis absorbance measured at 545 nm. The results of each experiment were normalized to the results for a Triton X-100-lysed control. n = 4 biological replicates. Statistical differences were analyzed with two-way ANOVA and the Sidak multiple-comparison test. Significance thresholds: *, P < 0.05; ****, P < 0.0001; ns, not statistically significant (P > 0.05).
FIG 4 Pore formation is disrupted by mismatch between PopB and PcrV. (A, B) A549 cells were infected in the presence of propidium iodide with RP11946 (PAO1 ΔexsE ΔexoSTY pcrV+ ΔpcrH-popBD) or RP11948 [PAO1 ΔexsE ΔexoSTY pcrV::acrV(tip) ΔpcrH-popBD] producing PopD and the indicated translocators. RP2317 (PAO1 ΔexsE ΔexoSTY pcrV+) was the wild-type control. After 2 h, cells were washed and imaged (bright field and fluorescence with Texas red filter). (A) Representative micrographs from one biological replicate are shown. (B) Four biological replicates are quantified. Fluorescent cells were scored manually, and the results of each experiment were normalized to the results for the PopB/PcrV strain. Error bars show standard deviations. Statistical differences were analyzed with two-way ANOVA and the Sidak multiple-comparison test: *, P < 0.05;ns, not statistically significant (P > 0.05). (C) Sheep erythrocytes were pretreated with papain, which cleaves surface glycoproteins and exposes cryptic binding sites for P. aeruginosa hemagglutinin (53). The erythrocytes were infected for 1 h with strain RP3670 [PAO1 ΔexsE exoS(G/A−) pcrV+ ΔpcrH-popBD] or strain RP11222 [PAO1 ΔexsE exoS(G/A−) pcrV::Atip ΔpcrH-popBD] producing PopD and the indicated translocators. The erythrocyte suspension was then mixed, unbroken erythrocytes pelleted, and supernatant UV-Vis absorbance measured at 545 nm. The results of each experiment were normalized to the results for a Triton X-100-lysed control. n = 4 biological replicates. Statistical differences were analyzed with two-way ANOVA and the Sidak multiple-comparison test. Significance thresholds: *, P < 0.05; ****, P < 0.0001; ns, not statistically significant (P > 0.05).CC BY · PubMed Central
FIG 5 Deletion of the secretion regulator Pcr1 does not restore translocation for the translocator-mismatched (Bmix-PcrV) strain. A549 cells were infected for 2 h with strain RP3670 [PAO1 ΔexsE exoS(G/A−) pcrV+ ΔpcrH-popBD], strain RP11222 [PAO1 ΔexsE exoS(G/A−) pcrV::Atip ΔpcrH-popBD], strain RP6370 [PAO1 ΔexsE exoS(G/A−) pcrV+ ΔpcrH-popBD Δpcr1], or strain RP11226 [PAO1 ΔexsE exoS(G/A−) pcrV::Atip ΔpcrH-popBD Δpcr1] complemented with popD and the indicated B-translocator, along with both translocator chaperones, pcrH and acrH. The amount of translocated ExoS was assessed by lysing the host cells with either Triton X-100, which lyses the eukaryotic cell membrane but does not lyse the bacteria, or lysing with SDS, which lyses both the eukaryotic cell and peripherally attached bacteria. ExoS was also monitored in supernatant samples and in bacteria collected from the supernatant after infection (shown in Fig. S7). The presence of ExoS, tubulin (host cell cytoplasmic content), and RNA polymerase subunit α (RPOA, bacterial cytoplasmic content) was assessed by Western blotting. (A and C) Western blots of pcr1+ strains (A) and Δpcr1 strains (C). The amount of ExoS translocated into A549 cells is shown by the Triton-lysed samples. The lower-molecular-weight band is attributable to intracellular cleavage of ExoS (59). The positions of molecular-weight size markers (in kilodaltons) are indicated. SDS-lysed samples show the total amount of ExoS both translocated into host cells and remaining in attached bacteria. (B and D) Quantification of ExoS translocation in Triton-lysed samples for pcr1+ strains (B) and Δpcr1 strains (D). The amount of translocated ExoS is normalized to the amount of tubulin in the same strain and to the amount of ExoS in the strain producing PopB/PcrV. n = 6 biological replicates. Error bars show standard deviations. Statistical differences were analyzed with two-way ANOVA and the Sidak multiple-comparison test. Significance thresholds: *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ns, not statistically significant (P > 0.05).
FIG 5 Deletion of the secretion regulator Pcr1 does not restore translocation for the translocator-mismatched (Bmix-PcrV) strain. A549 cells were infected for 2 h with strain RP3670 [PAO1 ΔexsE exoS(G/A−) pcrV+ ΔpcrH-popBD], strain RP11222 [PAO1 ΔexsE exoS(G/A−) pcrV::Atip ΔpcrH-popBD], strain RP6370 [PAO1 ΔexsE exoS(G/A−) pcrV+ ΔpcrH-popBD Δpcr1], or strain RP11226 [PAO1 ΔexsE exoS(G/A−) pcrV::Atip ΔpcrH-popBD Δpcr1] complemented with popD and the indicated B-translocator, along with both translocator chaperones, pcrH and acrH. The amount of translocated ExoS was assessed by lysing the host cells with either Triton X-100, which lyses the eukaryotic cell membrane but does not lyse the bacteria, or lysing with SDS, which lyses both the eukaryotic cell and peripherally attached bacteria. ExoS was also monitored in supernatant samples and in bacteria collected from the supernatant after infection (shown in Fig. S7). The presence of ExoS, tubulin (host cell cytoplasmic content), and RNA polymerase subunit α (RPOA, bacterial cytoplasmic content) was assessed by Western blotting. (A and C) Western blots of pcr1+ strains (A) and Δpcr1 strains (C). The amount of ExoS translocated into A549 cells is shown by the Triton-lysed samples. The lower-molecular-weight band is attributable to intracellular cleavage of ExoS (59). The positions of molecular-weight size markers (in kilodaltons) are indicated. SDS-lysed samples show the total amount of ExoS both translocated into host cells and remaining in attached bacteria. (B and D) Quantification of ExoS translocation in Triton-lysed samples for pcr1+ strains (B) and Δpcr1 strains (D). The amount of translocated ExoS is normalized to the amount of tubulin in the same strain and to the amount of ExoS in the strain producing PopB/PcrV. n = 6 biological replicates. Error bars show standard deviations. Statistical differences were analyzed with two-way ANOVA and the Sidak multiple-comparison test. Significance thresholds: *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ns, not statistically significant (P > 0.05).CC BY · PubMed Central
FIG 6 Models of PcrV-assisted translocation pore formation. (A) Tether model: PcrV prevents lateral diffusion of PopB and PopD in the membrane, encouraging oligomerization. (B) Scaffold model: the pore proteins could oligomerize around the PcrV needle tip as they are secreted. (C) Prepore model: PopB and PopD could form a prepore structure which is converted by PcrV to its final form. Schematic figures were created using BioRender.com.
FIG 6 Models of PcrV-assisted translocation pore formation. (A) Tether model: PcrV prevents lateral diffusion of PopB and PopD in the membrane, encouraging oligomerization. (B) Scaffold model: the pore proteins could oligomerize around the PcrV needle tip as they are secreted. (C) Prepore model: PopB and PopD could form a prepore structure which is converted by PcrV to its final form. Schematic figures were created using BioRender.com.CC BY · PubMed Central

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Metadata

Authors
Kundracik, E. C., Trichka, J., Díaz Aponte, J. E., & Rietsch, A.
Affiliations
  • Case Western Reserve University (US)
  • National Institutes of Health (US)
Corresponding author
Rietsch, Arne
ORCID
Venue
mBio
Publisher
openRxiv
Published
2022-01-22
Volume
13
Issue
5
Pages
e0238122-e0238122
ISSN
2150-7511 (ISSN-L)
DOI
10.1101/2022.01.21.477233
PMC
PMC9600203
OpenAlex
W4297220006
Cited by
18
References
1
Citations by year
2026: 5 · 2025: 7 · 2024: 1 · 2023: 4 · 2022: 1
FWCI
2.05
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • mBio (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Case Western Reserve University
  • Imperial College London
  • Universitat de Barcelona
  • National Institutes of Health (GM007250)
  • National Eye Institute (R01EY022052, P30EY011373)
  • National Institute of General Medical Sciences (T32GM007250)
  • National Institute of Allergy and Infectious Diseases (R21AI107131, R21AI168028)
Topics
  • Life Sciences › Immunology and Microbiology › Microbiology › Escherichia coli research studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Pathogenic Bacteria Studies
Keywords
type III secretion system, translocon, PopB, PcrV, pore formation, protein-protein interaction, needle tip, Pseudomonas aeruginosa, effector delivery, host-pathogen interface
MeSH terms
Type III Secretion Systems, Antigens, Bacterial, Bacterial Proteins, Pseudomonas aeruginosa, Virulence Factors

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.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: PopB-PcrV Interactions are Essential for Pore Formation in the Pseudomonas aeruginosa Type III Secretion System Translocon. https://app.cochat.ai/papers/10.1101/2022.01.21.477233
The paper
Kundracik, E. C., Trichka, J., Díaz Aponte, J. E., & Rietsch, A. (2022). PopB-PcrV Interactions are Essential for Pore Formation in the Pseudomonas aeruginosa Type III Secretion System Translocon. mBio. https://doi.org/10.1101/2022.01.21.477233

Data sources

Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.