Student or educator? Sign up with your school email and get 6 months of Pro free.
CoChat
Sign Up
Cited by 2Nuclear Structure and FunctionDNA Repair MechanismsMicrotubule and mitosis dynamics

The DOI matches its Crossref record; 91 of the 91 references it cites were found in a registry or an index.

Exportin-mediated nucleocytoplasmic transport maintains Pch2 homeostasis during meiosis

Open access
Herruzo, Esther, Sánchez-Díaz, Estefanía, González-Arranz, Sara, Santos, Beatriz, Carballo, Jesús A., San-Segundo, Pedro A.·2023·PLOS Genetics
DOI 10.1371/journal.pgen.1011026PMC PMC10688877

Published version of a preprint

Open this paper in CoChat — free accountOpen PDF

Abstract

The meiotic recombination checkpoint reinforces the order of events during meiotic prophase I, ensuring the accurate distribution of chromosomes to the gametes. The AAA+ ATPase Pch2 remodels the Hop1 axial protein enabling adequate levels of Hop1-T318 phosphorylation to support the ensuing checkpoint response. While these events are localized at chromosome axes, the checkpoint activating function of Pch2 relies on its cytoplasmic population. In contrast, forced nuclear accumulation of Pch2 leads to checkpoint inactivation. Here, we reveal the mechanism by which Pch2 travels from the cell nucleus to the cytoplasm to maintain Pch2 cellular homeostasis. Leptomycin B treatment provokes the nuclear accumulation of Pch2, indicating that its nucleocytoplasmic transport is mediated by the Crm1 exportin recognizing proteins containing Nuclear Export Signals (NESs). Consistently, leptomycin B leads to checkpoint inactivation and impaired Hop1 axial localization. Pch2 nucleocytoplasmic traffic is independent of its association with Zip1 and Orc1. We also identify a functional NES in the non-catalytic N-terminal domain of Pch2 that is required for its nucleocytoplasmic trafficking and proper checkpoint activity. In sum, we unveil another layer of control of Pch2 function during meiosis involving nuclear export via the exportin pathway that is crucial to maintain the critical balance of Pch2 distribution among different cellular compartments.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pgen.1011026

Cites

91 of 91 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig 1 Nuclear export of Pch2 is blocked by leptomycin B (LMB). (A) Schematic representation of protein nuclear export via nuclear pore complexes mediated by the interaction of a Nuclear Export Signal (NES) with the Crm1 exportin (see text for details). Leptomycin B (LMB) blocks NES binding to Crm1 preventing nuclear export. The presence of a putative NES in Pch2 is depicted. For simplicity, the NES is drawn only in one subunit of the Pch2 hexamer. BioRender.com was used to create this figure. (B, C) Fluorescence microscopy images of GFP-Pch2 localization in ZIP1 and zip1Δ cells, mock-treated (B), or treated with 500 ng/ml LMB (C), 15 h after meiotic induction. Images were taken at 24 h. Two representative individual cells for each condition are shown. Localization of GFP-Pch2 in the nucleolus (nlo), chromosomes (chr), and cytoplasm (cyt) is indicated. Scale bar, 2 μm. Strains are: DP1927 (ndt80Δ crm1-T539C GFP-PCH2) and DP1837 (zip1Δ ndt80Δ crm1-T539C GFP-PCH2). (D) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (B, C). Error bars, SD. (E) Western blot analysis of GFP-Pch2 production (using anti-Pch2 antibodies) in the absence (mock) or presence of LMB. Pgk1 was used as loading control. The graph shows the quantification of GFP-Pch2 relative levels. (F, G) Immunofluorescence of spread meiotic chromosomes at pachytene stained with anti-Pch2 antibodies (to detect GFP-Pch2; green), anti-Nsr1 antibodies (red) and DAPI (blue). Representative ZIP1 (F) and zip1Δ (G) nuclei, mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction are shown. Arrows point to the rDNA region. Spreads were prepared at 19 h. Scale bar, 2 μm. Strains are: DP1717 (crm1-T539C GFP-PCH2) and DP1721 (zip1Δ crm1-T539C GFP-PCH2). (H) Quantification of the GFP-Pch2 total signal for the experiment shown in (F, G). Error bars, SD; a.u., arbitrary units. (I-J) Quantification of the nucleolar (I) and chromosomal (J) GFP-Pch2 signal for the experiment shown in (F). Error bars, SD; a.u., arbitrary units.
Fig 1 Nuclear export of Pch2 is blocked by leptomycin B (LMB). (A) Schematic representation of protein nuclear export via nuclear pore complexes mediated by the interaction of a Nuclear Export Signal (NES) with the Crm1 exportin (see text for details). Leptomycin B (LMB) blocks NES binding to Crm1 preventing nuclear export. The presence of a putative NES in Pch2 is depicted. For simplicity, the NES is drawn only in one subunit of the Pch2 hexamer. BioRender.com was used to create this figure. (B, C) Fluorescence microscopy images of GFP-Pch2 localization in ZIP1 and zip1Δ cells, mock-treated (B), or treated with 500 ng/ml LMB (C), 15 h after meiotic induction. Images were taken at 24 h. Two representative individual cells for each condition are shown. Localization of GFP-Pch2 in the nucleolus (nlo), chromosomes (chr), and cytoplasm (cyt) is indicated. Scale bar, 2 μm. Strains are: DP1927 (ndt80Δ crm1-T539C GFP-PCH2) and DP1837 (zip1Δ ndt80Δ crm1-T539C GFP-PCH2). (D) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (B, C). Error bars, SD. (E) Western blot analysis of GFP-Pch2 production (using anti-Pch2 antibodies) in the absence (mock) or presence of LMB. Pgk1 was used as loading control. The graph shows the quantification of GFP-Pch2 relative levels. (F, G) Immunofluorescence of spread meiotic chromosomes at pachytene stained with anti-Pch2 antibodies (to detect GFP-Pch2; green), anti-Nsr1 antibodies (red) and DAPI (blue). Representative ZIP1 (F) and zip1Δ (G) nuclei, mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction are shown. Arrows point to the rDNA region. Spreads were prepared at 19 h. Scale bar, 2 μm. Strains are: DP1717 (crm1-T539C GFP-PCH2) and DP1721 (zip1Δ crm1-T539C GFP-PCH2). (H) Quantification of the GFP-Pch2 total signal for the experiment shown in (F, G). Error bars, SD; a.u., arbitrary units. (I-J) Quantification of the nucleolar (I) and chromosomal (J) GFP-Pch2 signal for the experiment shown in (F). Error bars, SD; a.u., arbitrary units.CC BY · PubMed Central
Fig 2 LMB treatment impairs the meiotic recombination checkpoint. (A) Immunofluorescence of zip1Δ spread meiotic chromosomes stained with anti-Hop1 (red) and anti-GFP (to detect GFP-Pch2; green) antibodies. Representative nuclei, either mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction, are shown. Arrows point to the rDNA region. Spreads were prepared at 19 h. Scale bar, 2 μm. The strain is DP1721 (zip1Δ crm1-T539C GFP-PCH2). (B) Quantification of Pch2 (left graph) and Hop1 (right graph) signal for the experiment shown in (A). Error bars, SD; a.u., arbitrary units. (C) Western blot analysis of Hop1-T318 phosphorylation (ph) in the absence (mock) or presence of LMB added at 15 h after meiotic induction (arrows). Pgk1 was used as loading control. The graph shows the quantification of Hop1-T318ph relative levels. Strains are: DP1927 (ndt80Δ crm1-T539C GFP-PCH2) and DP1837 (zip1Δ ndt80Δ crm1-T539C GFP-PCH2).
Fig 2 LMB treatment impairs the meiotic recombination checkpoint. (A) Immunofluorescence of zip1Δ spread meiotic chromosomes stained with anti-Hop1 (red) and anti-GFP (to detect GFP-Pch2; green) antibodies. Representative nuclei, either mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction, are shown. Arrows point to the rDNA region. Spreads were prepared at 19 h. Scale bar, 2 μm. The strain is DP1721 (zip1Δ crm1-T539C GFP-PCH2). (B) Quantification of Pch2 (left graph) and Hop1 (right graph) signal for the experiment shown in (A). Error bars, SD; a.u., arbitrary units. (C) Western blot analysis of Hop1-T318 phosphorylation (ph) in the absence (mock) or presence of LMB added at 15 h after meiotic induction (arrows). Pgk1 was used as loading control. The graph shows the quantification of Hop1-T318ph relative levels. Strains are: DP1927 (ndt80Δ crm1-T539C GFP-PCH2) and DP1837 (zip1Δ ndt80Δ crm1-T539C GFP-PCH2).CC BY · PubMed Central
Fig 3 Pch2 nucleocytoplasmic traffic does not involve its association with Orc1 and Zip1. (A) Fluorescence microscopy images of GFP-Pch2 localization in ZIP1 orc1-3mAID and zip1Δ orc1-3mAID cells, mock-treated, or treated with 500 ng/ml LMB, 15 h after meiotic induction. Auxin (500μM) was added to the cultures 12 h after meiotic induction to induce Orc1 degradation. Images were taken at 19 h. Cells representing the predominant GFP-Pch2 localization pattern for each condition are shown. Additional examples of GFP-Pch2 distribution are presented in S2D Fig. Scale bar, 2 μm. Strains are: DP1885 (orc1-3mAID crm1-T539C GFP-PCH2) and DP1886 (zip1Δ orc1-3mAID crm1-T539C GFP-PCH2). (B) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (A). Error bars, SD. (C-D) Immunofluorescence of ZIP1 orc1-3mAID (C) and zip1Δ orc1-3mAID (D) spread meiotic chromosomes stained with anti-Pch2 (green) and anti-Nsr1 (red) antibodies, and DAPI (blue). Representative nuclei, either mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction, are shown. Auxin (500 μM) was added to the cultures 12 h after meiotic induction to induce Orc1 degradation. Spreads were prepared at 19 h. Arrows point to the nucleolar region (nlo) and the Polycomplex (PC). Scale bar, 2 μm. The strains are the same used in (A). (E) Quantification of Pch2 signal for the experiment shown in (C-D). Error bars, SD; a.u., arbitrary units.
Fig 3 Pch2 nucleocytoplasmic traffic does not involve its association with Orc1 and Zip1. (A) Fluorescence microscopy images of GFP-Pch2 localization in ZIP1 orc1-3mAID and zip1Δ orc1-3mAID cells, mock-treated, or treated with 500 ng/ml LMB, 15 h after meiotic induction. Auxin (500μM) was added to the cultures 12 h after meiotic induction to induce Orc1 degradation. Images were taken at 19 h. Cells representing the predominant GFP-Pch2 localization pattern for each condition are shown. Additional examples of GFP-Pch2 distribution are presented in S2D Fig. Scale bar, 2 μm. Strains are: DP1885 (orc1-3mAID crm1-T539C GFP-PCH2) and DP1886 (zip1Δ orc1-3mAID crm1-T539C GFP-PCH2). (B) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (A). Error bars, SD. (C-D) Immunofluorescence of ZIP1 orc1-3mAID (C) and zip1Δ orc1-3mAID (D) spread meiotic chromosomes stained with anti-Pch2 (green) and anti-Nsr1 (red) antibodies, and DAPI (blue). Representative nuclei, either mock-treated, or treated with 500 ng/ml LMB 15 h after meiotic induction, are shown. Auxin (500 μM) was added to the cultures 12 h after meiotic induction to induce Orc1 degradation. Spreads were prepared at 19 h. Arrows point to the nucleolar region (nlo) and the Polycomplex (PC). Scale bar, 2 μm. The strains are the same used in (A). (E) Quantification of Pch2 signal for the experiment shown in (C-D). Error bars, SD; a.u., arbitrary units.CC BY · PubMed Central
Fig 4 A functional NES in the NTD of Pch2 promotes its nuclear export. (A) Fluorescence microscopy images of the localization of GFP-Pch2, GFP-Pch2-nes4A and GFP-NESPKI-Pch2-nes4A in zip1Δ cells. Images were taken 15 h after meiotic induction. Representative cells are shown. A schematic representation of the different GFP-Pch2 versions is also shown on top. Scale bar, 2 μm. Strains are: DP1625 (zip1Δ GFP-PCH2), DP1986 (zip1Δ GFP-pch2-nes4A) and DP1992 (zip1Δ GFP-NESPKI-pch2-nes4A). (B) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (A). Error bars, SD. (C) Time course analysis of meiotic nuclear divisions; the percentage of cells containing two or more nuclei is represented. Error bars: SD; n = 3. At least 300 cells were scored for each strain at every time point. Strains are: DP1624 (GFP-PCH2), DP1625 (zip1Δ GFP-PCH2), DP1029 (zip1Δ pch2Δ), DP1986 (zip1Δ GFP-pch2-nes4A) and DP1992 (zip1Δ GFP-NESPKI-pch2-nes4A). (D) Immunofluorescence of spread meiotic chromosomes stained with anti-Hop1 antibodies and DAPI (blue). Representative nuclei from prophase-arrested ndt80Δ strains are shown. Arrows point to the rDNA region. Spreads were prepared at 24 h. Scale bar, 2 μm. Strains are: DP1655 (ndt80Δ zip1Δ GFP-PCH2), DP1988 (ndt80Δ zip1Δ GFP-pch2-nes4A), DP2003 (ndt80Δ zip1Δ GFP-NESPKI-pch2-nes4A), and DP881 (ndt80Δ zip1Δ pch2Δ). (E) Quantification of the Hop1 signal for the experiment shown in (D). (F) Western blot analysis of Hop1-T318 phosphorylation in the strains analyzed in (C) at the indicated time points in meiosis. Pgk1 was used as a loading control. The graph shows the quantification of Hop1-T318ph relative levels.
Fig 4 A functional NES in the NTD of Pch2 promotes its nuclear export. (A) Fluorescence microscopy images of the localization of GFP-Pch2, GFP-Pch2-nes4A and GFP-NESPKI-Pch2-nes4A in zip1Δ cells. Images were taken 15 h after meiotic induction. Representative cells are shown. A schematic representation of the different GFP-Pch2 versions is also shown on top. Scale bar, 2 μm. Strains are: DP1625 (zip1Δ GFP-PCH2), DP1986 (zip1Δ GFP-pch2-nes4A) and DP1992 (zip1Δ GFP-NESPKI-pch2-nes4A). (B) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (A). Error bars, SD. (C) Time course analysis of meiotic nuclear divisions; the percentage of cells containing two or more nuclei is represented. Error bars: SD; n = 3. At least 300 cells were scored for each strain at every time point. Strains are: DP1624 (GFP-PCH2), DP1625 (zip1Δ GFP-PCH2), DP1029 (zip1Δ pch2Δ), DP1986 (zip1Δ GFP-pch2-nes4A) and DP1992 (zip1Δ GFP-NESPKI-pch2-nes4A). (D) Immunofluorescence of spread meiotic chromosomes stained with anti-Hop1 antibodies and DAPI (blue). Representative nuclei from prophase-arrested ndt80Δ strains are shown. Arrows point to the rDNA region. Spreads were prepared at 24 h. Scale bar, 2 μm. Strains are: DP1655 (ndt80Δ zip1Δ GFP-PCH2), DP1988 (ndt80Δ zip1Δ GFP-pch2-nes4A), DP2003 (ndt80Δ zip1Δ GFP-NESPKI-pch2-nes4A), and DP881 (ndt80Δ zip1Δ pch2Δ). (E) Quantification of the Hop1 signal for the experiment shown in (D). (F) Western blot analysis of Hop1-T318 phosphorylation in the strains analyzed in (C) at the indicated time points in meiosis. Pgk1 was used as a loading control. The graph shows the quantification of Hop1-T318ph relative levels.CC BY · PubMed Central
Fig 5 The pch2-nes4A mutant has little impact on an unperturbed ZIP1 meiosis. (A) Time course analysis of meiotic nuclear divisions; the percentage of cells containing two or more nuclei is represented. Error bars: SD; n = 3. At least 300 cells were scored for each strain at every time point. (B) Sporulation efficiency, assessed by microscopic counting of asci, was examined after 3 days on sporulation plates. Error bars, SD; n = 3. At least 600 cells were counted for each strain. (C) Spore viability, assessed by tetrad dissection, is shown in the left graph. The percentage of tetrads containing 4-, 3-, 2-, 1-, and 0-viable spores is presented in the right graph. At least 144 tetrads were dissected for each strain. Error bars, SD. (D, E) Immunofluorescence of spread meiotic chromosomes at pachytene stained with anti-GFP antibodies (to detect GFP-Pch2; green) and anti-Zip1 antibodies (red), and DAPI (blue). Representative nuclei are shown. Spreads were prepared 15 h (D) or 24 h (E) after meiotic induction. Arrows point to the rDNA region (nlo) and polycomplex (PC). Scale bar, 2 μm. (F) Quantification of Pch2 signal on spread nuclei from the experiment shown in (D, E). a.u., arbitrary units. (G) Percentage of nuclei containing polycomplexes in the experiment shown in (D, E). Between 23 to 30 nuclei were scored in (F, G). Strains are: DP1023 (pch2Δ), DP1620 (GFP-PCH2), DP2052 (GFP-pch2-nes4A), DP1788 (spo11-3HA GFP-PCH2), DP2046 (spo11-3HA GFP-pch2-nes4A), DP1787 (spo11-3HA pch2Δ), DP1639 (ndt80Δ GFP-PCH2), and DP2053 (ndt80Δ GFP-pch2-nes4A).
Fig 5 The pch2-nes4A mutant has little impact on an unperturbed ZIP1 meiosis. (A) Time course analysis of meiotic nuclear divisions; the percentage of cells containing two or more nuclei is represented. Error bars: SD; n = 3. At least 300 cells were scored for each strain at every time point. (B) Sporulation efficiency, assessed by microscopic counting of asci, was examined after 3 days on sporulation plates. Error bars, SD; n = 3. At least 600 cells were counted for each strain. (C) Spore viability, assessed by tetrad dissection, is shown in the left graph. The percentage of tetrads containing 4-, 3-, 2-, 1-, and 0-viable spores is presented in the right graph. At least 144 tetrads were dissected for each strain. Error bars, SD. (D, E) Immunofluorescence of spread meiotic chromosomes at pachytene stained with anti-GFP antibodies (to detect GFP-Pch2; green) and anti-Zip1 antibodies (red), and DAPI (blue). Representative nuclei are shown. Spreads were prepared 15 h (D) or 24 h (E) after meiotic induction. Arrows point to the rDNA region (nlo) and polycomplex (PC). Scale bar, 2 μm. (F) Quantification of Pch2 signal on spread nuclei from the experiment shown in (D, E). a.u., arbitrary units. (G) Percentage of nuclei containing polycomplexes in the experiment shown in (D, E). Between 23 to 30 nuclei were scored in (F, G). Strains are: DP1023 (pch2Δ), DP1620 (GFP-PCH2), DP2052 (GFP-pch2-nes4A), DP1788 (spo11-3HA GFP-PCH2), DP2046 (spo11-3HA GFP-pch2-nes4A), DP1787 (spo11-3HA pch2Δ), DP1639 (ndt80Δ GFP-PCH2), and DP2053 (ndt80Δ GFP-pch2-nes4A).CC BY · PubMed Central
Fig 6 Fusion of a putative NES from human TRIP13 restores checkpoint function to Pch2-nes4A. (A) Schematic representation of the S. cerevisiae Pch2 protein (ScPch2) and the human ortholog (HsTRIP13) indicating the characteristic AAA+ ATPase motifs, the conserved N-terminal domain (NTD) and the extended N-terminal domain exclusive of yeast Pch2 (NTDEXT). The positions and sequences of the putative NESs analyzed in this work are depicted (purple boxes). (B) Fluorescence microscopy images of the localization of GFP-Pch2, GFP-Pch2-nes4A, GFP-NESTRIP13-Pch2-nes4A, and GFP-nes7ATRIP13-Pch2-nes4A in zip1Δ cells. Images were taken 16 h after meiotic induction. Representative cells are shown. A schematic representation of the different GFP-Pch2 versions is also shown on top. Scale bar, 2 μm. The strain is DP1405 (zip1Δ pch2Δ) transformed with the centromeric plasmids pSS393 (GFP-PCH2), pSS459 (GFP-pch2-nes4A), pSS472 (GFP-NESTRIP13-pch2-nes4A) and pSS474 (GFP-nes7ATRIP13-pch2-nes4A). (C) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (B). Error bars, SD. (D) Time course analysis of meiotic nuclear divisions. The percentage of cells containing two or more nuclei is represented. Error bars: SD; n = 3. At least 300 cells were scored for each strain at every time point. Strains are: DP1625 (zip1Δ GFP-PCH2), DP1986 (zip1Δ GFP-pch2-nes4A), DP2025 (zip1Δ GFP-NESTRIP13-pch2-nes4A) and DP2033 (zip1Δ GFP-nes7ATRIP13-pch2-nes4A). (E) Western blot analysis of Hop1-T318 phosphorylation in the strains analyzed in (D) at the indicated time points in meiosis. Pgk1 was used as a loading control. The graph shows the quantification of Hop1-T318ph relative levels. (F) Pch2 structure modeled by AlphaFold. (G) Human TRIP13E253Q structure obtained from the Protein Data Bank (PDB 5VQ9) [42]. (H) AlphaFold predicted structure resulting from the fusion of NESTRIP13 to Pch2-nes4A. In (F-H), the N-terminal domains of Pch2 and TRIP13 are labeled in pink, the position of NESPch2 (or nesPch2-4A) is labeled in blue, and the position of the putative NESTRIP13 is labeled in orange.
Fig 6 Fusion of a putative NES from human TRIP13 restores checkpoint function to Pch2-nes4A. (A) Schematic representation of the S. cerevisiae Pch2 protein (ScPch2) and the human ortholog (HsTRIP13) indicating the characteristic AAA+ ATPase motifs, the conserved N-terminal domain (NTD) and the extended N-terminal domain exclusive of yeast Pch2 (NTDEXT). The positions and sequences of the putative NESs analyzed in this work are depicted (purple boxes). (B) Fluorescence microscopy images of the localization of GFP-Pch2, GFP-Pch2-nes4A, GFP-NESTRIP13-Pch2-nes4A, and GFP-nes7ATRIP13-Pch2-nes4A in zip1Δ cells. Images were taken 16 h after meiotic induction. Representative cells are shown. A schematic representation of the different GFP-Pch2 versions is also shown on top. Scale bar, 2 μm. The strain is DP1405 (zip1Δ pch2Δ) transformed with the centromeric plasmids pSS393 (GFP-PCH2), pSS459 (GFP-pch2-nes4A), pSS472 (GFP-NESTRIP13-pch2-nes4A) and pSS474 (GFP-nes7ATRIP13-pch2-nes4A). (C) Quantification of the ratio of nuclear (including nucleolar) to cytoplasmic GFP fluorescent signal for the experiment shown in (B). Error bars, SD. (D) Time course analysis of meiotic nuclear divisions. The percentage of cells containing two or more nuclei is represented. Error bars: SD; n = 3. At least 300 cells were scored for each strain at every time point. Strains are: DP1625 (zip1Δ GFP-PCH2), DP1986 (zip1Δ GFP-pch2-nes4A), DP2025 (zip1Δ GFP-NESTRIP13-pch2-nes4A) and DP2033 (zip1Δ GFP-nes7ATRIP13-pch2-nes4A). (E) Western blot analysis of Hop1-T318 phosphorylation in the strains analyzed in (D) at the indicated time points in meiosis. Pgk1 was used as a loading control. The graph shows the quantification of Hop1-T318ph relative levels. (F) Pch2 structure modeled by AlphaFold. (G) Human TRIP13E253Q structure obtained from the Protein Data Bank (PDB 5VQ9) [42]. (H) AlphaFold predicted structure resulting from the fusion of NESTRIP13 to Pch2-nes4A. In (F-H), the N-terminal domains of Pch2 and TRIP13 are labeled in pink, the position of NESPch2 (or nesPch2-4A) is labeled in blue, and the position of the putative NESTRIP13 is labeled in orange.CC BY · PubMed Central
Fig 7 NES-mediated nuclear export of Pch2 is essential for meiotic recombination checkpoint function. The nucleocytoplasmic transport of Pch2 via the exportin pathway maintains Pch2 homeostasis and supports proper checkpoint activity in zip1Δ. In the NES-deficient pch2-nes4A mutant, the accumulation of Pch2 in the nucleus leads to checkpoint inactivation. The fusion to Pch2-nes4A of a characterized ectopic NES from the PKI protein, or a putative NES from the human TRIP13 ortholog, restores the balanced subcellular distribution of Pch2 and restores checkpoint function. For simplicity, the NES is drawn only in one subunit of the Pch2 hexamer. BioRender.com was used to create this figure.
Fig 7 NES-mediated nuclear export of Pch2 is essential for meiotic recombination checkpoint function. The nucleocytoplasmic transport of Pch2 via the exportin pathway maintains Pch2 homeostasis and supports proper checkpoint activity in zip1Δ. In the NES-deficient pch2-nes4A mutant, the accumulation of Pch2 in the nucleus leads to checkpoint inactivation. The fusion to Pch2-nes4A of a characterized ectopic NES from the PKI protein, or a putative NES from the human TRIP13 ortholog, restores the balanced subcellular distribution of Pch2 and restores checkpoint function. For simplicity, the NES is drawn only in one subunit of the Pch2 hexamer. BioRender.com was used to create this figure.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 1

  1. K Mittal, 2022, Targeting TRIP13 in Wilms tumor with nuclear export inhibitors., bioRxiv, 2022.02.23.481521

Metadata

Authors
Herruzo, E., Sánchez-Díaz, E., González-Arranz, S., Santos, B., Carballo, J. A., & San-Segundo, P. A.
Affiliations
Consejo Superior de Investigaciones Científicas (ES)
Corresponding author
San-Segundo, Pedro A.
ORCID
Venue
PLOS Genetics
Publisher
Public Library of Science (PLoS)
Published
2023-11-10
Volume
19
Issue
11
Pages
e1011026
ISSN
  • 1553-7390 (ISSN-L)
  • 1553-7404
DOI
10.1371/journal.pgen.1011026
PMC
PMC10688877
OpenAlex
W4388562972
Cited by
2
References
91
Citations by year
2026: 1 · 2024: 1
FWCI
0.18
Type
journal article
Language
en
Update notices
new_version (2023-11-30)
Open access
Licence
Locations
  • PLoS Genetics (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DIGITAL.CSIC (Spanish National Research Council (CSIC)) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Ministerio de Ciencia e Innovación (PID2021-125830NB-I00)
  • Consejería de Educación, Junta de Castilla y León (CSI259P20)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
Keywords
Pch2, nuclear transport, nuclear export signal, AAA+ ATPase, leptomycin B, nuclear accumulation, meiotic prophase I, nuclear export
MeSH terms
DNA-Binding Proteins, Homeostasis, Meiosis, Nuclear Proteins, Saccharomyces cerevisiae, Active Transport, Cell Nucleus, Karyopherins, Saccharomyces cerevisiae Proteins

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 ›

Are you an author, or do you see a problem with this page? Request a review

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.