Spo13 prevents premature cohesin cleavage during meiosis

Abstract

Meiosis produces gametes through two successive nuclear Background: divisions, meiosis I and meiosis II. In contrast to mitosis and meiosis II, where sister chromatids are segregated, during meiosis I, homologous chromosomes are segregated. This requires the monopolar attachment of sister kinetochores and the loss of cohesion from chromosome arms, but not centromeres, during meiosis I. The establishment of both sister kinetochore mono-orientation and cohesin protection rely on the budding yeast meiosis I-specific Spo13 protein, the functional homolog of fission yeast Moa1 and mouse MEIKIN.Here we investigate the effects of loss of on cohesion Methods: SPO13 during meiosis I using a live-cell imaging approach.Unlike wild type, cells lacking fail to maintain the Results:SPO13 meiosis-specific cohesin subunit, Rec8, at centromeres and segregate sister chromatids to opposite poles during anaphase I. We show that the cohesin-destabilizing factor, Wpl1, is not primarily responsible for the loss of cohesion during meiosis I. Instead, premature loss of centromeric cohesin during anaphase I in cells relies on separase-dependent spo13Δ cohesin cleavage. Further, cohesin loss in anaphase I cells is spo13Δ blocked by forcibly tethering the regulatory subunit of protein phosphatase 2A, Rts1, to Rec8.Our findings indicate that separase-dependent cleavage of Conclusions: phosphorylated Rec8 causes premature cohesin loss in cells. spo13Δ ( ) First published: 4 https://doi.org/10.12688/wellcomeopenres.15066.1 Argüello-Miranda O, Zagoriy I, Mengoli V, et al.: Casein Kinase 1 Coordinates Cohesin Cleavage, Gametogenesis, and Exit from M Phase in Meiosis II. Dev Cell. 2017; 40(1): 37-52. PubMed Abstract | Publisher Full Text Attner MA, Miller MP, Ee LS, et al.: Polo kinase Cdc5 is a central regulator of meiosis I. Proc Natl Acad Sci U S A. 2013; 110(35): 14278-14283. PubMed Abstract | Publisher Full Text | Free Full Text Rolef Ben-Shahar T, Heeger S, Lehane C, et al.: Eco1-Dependent Cohesin Acetylation During Establishment of Sister Chromatid Cohesion. Science. 2008; 321(5888): 563-566. PubMed Abstract | Publisher Full Text Brar GA, Kiburz BM, Zhang Y, et al.: Rec8 phosphorylation and recombination promote the step-wise loss of cohesins in meiosis. Nature. 2006; 441(7092): 532-536. PubMed Abstract | Publisher Full Text Buheitel J, Stemmann O: Prophase pathway-dependent removal of cohesin from human chromosomes requires opening of the Smc3-Scc1 gate. EMBO J. 2013; 32(5): 666-676. PubMed Abstract | Publisher Full Text | Free Full Text Buonomo SB, Clyne RK, Fuchs J, et al.: Disjunction of homologous chromosomes in meiosis I depends on proteolytic cleavage of the meiotic cohesin Rec8 by separin. Cell. 2000; 103(3): 387-398. PubMed Abstract | Publisher Full Text Challa K, Fajish VG, Shinohara M, et al.: Meiosis-specific prophase-like pathway controls cleavage-independent release of cohesin by Wapl phosphorylation. PLoS Genet. 2019; 15(1): e1007851. PubMed Abstract | Publisher Full Text | Free Full Text Challa K, Lee MS, Shinohara M, et al...

Reductional Meiosis I Chromosome Segregation Is Established by Coordination of Key Meiotic Kinases

“… The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository. This study PXD012627 Experimental Models: Organisms/Strains Yeast strains used in this study n/a See Table S1 Oligonucleotides Oligonucleotides used in this study for qPCR n/a See Table S3 Recombinant DNA Plasmids generated in this study n/a See Table S2 Software Image J plugin “DV_DotCounter” for analysis of microscopy data is available on Github This study and Galander et al., 2019 https://doi.org/10.5281/zenodo.2553082 ) Image J plugin “YeastLineProfiler” for analysis of microscopy data is available on Github This study and Galander et al., 2019 https://doi.org/10.5281/zenodo.2560099 Other NEXTflex-6 DNA Barcodes Perkin Elmer 514101 …”

“…An attractive candidate is the budding yeast meiosis-I-specific Spo13 protein. Cells lacking SPO13 undergo a single meiotic division, show monoorientation defects, and fail to protect cohesin ( Katis et al., 2004b , Klapholz and Esposito, 1980 , Lee et al., 2004 , Shonn et al., 2002 , Galander et al., 2019 ). Accordingly, Spo13 is required for monopolin localization at kinetochores ( Katis et al., 2004b , Lee et al., 2004 ) and is implicated in ensuring the proper pericentromeric localization of Sgo1 ( Kiburz et al., 2005 ).…”

Meiosis I Kinase Regulators: Conserved Orchestrators of Reductional Chromosome Segregation

“…[ 48–51 ] There, PP2A is thought to dephosphorylate cohesin, thus preventing its cleavage by separase. Artificial tethering of PP2A to arm cohesin in fission yeast [ 51 ] or budding yeast [ 52 ] impairs cohesin cleavage on chromosome arms, highlighting the importance of restricting PP2A activity to the pericentromere. Thus, cohesin protection in the pericentromere requires the careful balancing of cohesin‐directed kinase and phosphatase activity; disruption of this balance is likely to interfere with the maintenance of pericentromeric cohesin after metaphase I.…”

“…spo13Δ mutants, for example, only undergo a single meiotic division [ 82 ] characterized by a mixture of reductional and equational chromosome segregation. [ 52,53 ] This phenotype is not seen in fission yeast moa1Δ cells [ 10 ] or mouse Meikin −/− oocytes. [ 15 ] Little is known about the molecular basis of the altered cell cycle in spo13Δ cells, but it has been observed that deletion of the spindle checkpoint component MAD2 restores the second division in a majority of spo13Δ cells, [ 53 ] although chromosome segregation is still defective in spo13Δ mad2Δ strains.…”

Mechanism ofin vivoactivation of the MutLγ-Exo1 complex for meiotic crossover formation

“…including Cdc5, in pericentromeric regions (Galander et al, 2019). It may also 12 prevent Cdc5-driven MutLg activation in these regions.…”