Psm3 Acetylation on Conserved Lysine Residues Is Dispensable for Viability in Fission Yeast but Contributes to Eso1-Mediated Sister Chromatid Cohesion by Antagonizing Wpl1

Sister chromatid cohesion, which is established during the S phase of the eukaryotic cell cycle and persists until the onset of anaphase, is essential for the maintenance of genomic integrity. Cohesion requires the multi-protein complex cohesin, as well as a number of accessory proteins including Pds5/BIMD/Spo76. In the budding yeast Saccharomyces cerevisiae Pds5 is an essential protein that localises to chromosomes in a cohesin-dependent manner. Here we describe the characterisation in the fission yeast Schizosaccharomyces pombe of pds5+, a novel,non-essential orthologue of S. cerevisiae PDS5. The S. pombePds5 protein was localised to punctate nuclear foci in a manner that was dependent on the Rad21 cohesin component. This, together with additional genetic evidence, points towards an involvement of S. pombe Pds5 in sister chromatid cohesion. S. pombe pds5 mutants were hypersensitive to DNA damage and to mitotic metaphase delay, but this sensitivity was apparently not due to precocious loss of sister chromatid cohesion. These cells also suffered increased spontaneous chromosome loss and meiotic defects and their viability was dependent on the spindle checkpoint protein Bub1. Thus, while S. pombe Pds5 has an important cohesin-related role, this differs significantly from that of the equivalent budding yeast protein.

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Rec8p, a Meiotic Recombination and Sister Chromatid Cohesion Phosphoprotein of the Rad21p Family Conserved from Fission Yeast to Humans

Molecular and Cellular Biology ◽

10.1128/mcb.19.5.3515 ◽

1999 ◽

Vol 19 (5) ◽

pp. 3515-3528 ◽

Cited By ~ 144

Author(s):

Sandro Parisi ◽

Michael J. McKay ◽

Monika Molnar ◽

M. Anne Thompson ◽

Peter J. van der Spek ◽

...

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Germ Line ◽

Sequence Similarity ◽

Sister Chromatid Cohesion ◽

Specific Expression ◽

Chromatid Cohesion ◽

Prophase Nucleus ◽

Chromosome Iii ◽

Meiosis I

ABSTRACT Our work and that of others defined mitosis-specific (Rad21 subfamily) and meiosis-specific (Rec8 subfamily) proteins involved in sister chromatid cohesion in several eukaryotes, including humans. Mutation of the fission yeast Schizosaccharomyces pombe rec8 gene was previously shown to confer a number of meiotic phenotypes, including strong reduction of recombination frequencies in the central region of chromosome III, absence of linear element polymerization, reduced pairing of homologous chromosomes, reduced sister chromatid cohesion, aberrant chromosome segregation, defects in spore formation, and reduced spore viability. Here we extend the description of recombination reduction to the central regions of chromosomes I and II. We show at the protein level that expression ofrec8 is meiosis specific and that Rec8p localizes to approximately 100 foci per prophase nucleus. Rec8p was present in an unphosphorylated form early in meiotic prophase but was phosphorylated prior to meiosis I, as demonstrated by analysis of the mei4mutant blocked before meiosis I. Evidence for the persistence of Rec8p beyond meiosis I was obtained by analysis of the mutantmes1 blocked before meiosis II. A human gene, which we designate hrec8, showed significant primary sequence similarity to rec8 and was mapped to chromosome 14. High mRNA expression of mouse and human rec8 genes was found only in germ line cells, specifically in testes and, interestingly, in spermatids. hrec8 was also expressed at a low level in the thymus. Sequence similarity and testis-specific expression indicate evolutionarily conserved functions of Rec8p in meiosis. Possible roles of Rec8p in the integration of different meiotic events are discussed.

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The cohesin ring uses its hinge to organize DNA using non-topological as well as topological mechanisms

10.1101/197848 ◽

2017 ◽

Cited By ~ 2

Author(s):

Madhusudhan Srinivasan ◽

Johanna C. Scheinost ◽

Naomi J. Petela ◽

Thomas G. Gligoris ◽

Maria Wissler ◽

...

Keyword(s):

Conformational Changes ◽

Sister Chromatid ◽

Atp Hydrolysis ◽

Sister Chromatid Cohesion ◽

Perfect Correlation ◽

Chromatid Cohesion ◽

Lysine Residues ◽

Positively Charged ◽

Hinge Domain

SummaryAs predicted by the notion that sister chromatid cohesion is mediated by entrapment of sister DNAs inside cohesin rings, there is a perfect correlation between co-entrapment of circular minichromosomes and sister chromatid cohesion in a large variety of mutants. In most cells where cohesin loads onto chromosomes but fails to form cohesion, loading is accompanied by entrapment of individual DNAs. However, cohesin with a hinge domain whose positively charged lumen has been neutralized not only loads onto and translocates along chromatin but also organizes it into chromatid-like threads, despite largely failing to entrap DNAs inside its ring. Thus, cohesin engages chromatin in a non-topological as well as a topological manner. Our finding that hinge mutations, but not fusions between Smc and kleisin subunits, abolish entrapment suggests that DNAs may enter cohesin rings through hinge opening. Lastly, mutation of three highly conserved lysine residues inside the Smc1 moiety of Smc1/3 hinges abolishes all loading without affecting cohesin’s initial recruitment to CEN loading sites or its ability to hydrolyze ATP. We suggest that loading and translocation are mediated by conformational changes in cohesin’s hinge driven by cycles of ATP hydrolysis.

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Fission Yeast Eso1p Is Required for Establishing Sister Chromatid Cohesion during S Phase

Molecular and Cellular Biology ◽

10.1128/.20.10.3459-3469.2000 ◽

2000 ◽

Vol 20 (10) ◽

pp. 3459-3469 ◽

Cited By ~ 3

Author(s):

Koichi Tanaka ◽

Toshihiro Yonekawa ◽

Yosuke Kawasaki ◽

Mihoko Kai ◽

Kanji Furuya ◽

...

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

S Phase ◽

Chromatid Cohesion

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Establishment and maintenance of sister chromatid cohesion in fission yeast by a unique mechanism

The EMBO Journal ◽

10.1093/emboj/20.20.5779 ◽

2001 ◽

Vol 20 (20) ◽

pp. 5779-5790 ◽

Cited By ~ 109

Author(s):

K. Tanaka

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion ◽

Unique Mechanism

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Shugoshin protects cohesin complexes at centromeres

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2004.1607 ◽

2005 ◽

Vol 360 (1455) ◽

pp. 515-521 ◽

Cited By ~ 56

Author(s):

Yoshinori Watanabe ◽

Tomoya S Kitajima

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Crucial Role ◽

Regional Difference ◽

Sister Chromatid Cohesion ◽

Protein Family ◽

Chromatid Cohesion ◽

Spindle Microtubules ◽

Pulling Force ◽

Eukaryotic Organisms

The different regulation of sister chromatid cohesion at centromeres and along chromosome arms is obvious during meiosis, because centromeric cohesion, but not arm cohesion, persists throughout anaphase of the first division. A protein required to protect centromeric cohesin Rec8 from separase cleavage has been identified and named shugoshin (or Sgo1) after shugoshin (‘guardian spirit’ in Japanese). It has become apparent that shugoshin shows marginal homology with Drosophila Mei-S332 and several uncharacterized proteins in other eukaryotic organisms. Because Mei-S332 is a protein previously shown to be required for centromeric cohesion in meiosis, it is now established that shugoshin represents a conserved protein family defined as a centromeric protector of Rec8 cohesin complexes in meiosis. The regional difference of sister chromatid cohesion is also observed during mitosis in vertebrates; the cohesion is much more robust at the centromere at metaphase, where it antagonizes the pulling force of spindle microtubules that attach the kinetochores from opposite poles. The human shugoshin homologue (hSgo1) is required to protect the centromeric localization of the mitotic cohesin, Scc1, until metaphase. Bub1 plays a crucial role in the localization of shugoshin to centromeres in both fission yeast and humans.

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The acetyltransferase Eco1 elicits cohesin dimerization during S phase

Journal of Biological Chemistry ◽

10.1074/jbc.ra120.013102 ◽

2020 ◽

Vol 295 (22) ◽

pp. 7554-7565 ◽

Cited By ~ 3

Author(s):

Di Shi ◽

Shuaijun Zhao ◽

Mei-Qing Zuo ◽

Jingjing Zhang ◽

Wenya Hou ◽

...

Keyword(s):

Dna Replication ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

S Phase ◽

Cross Linking ◽

Temperature Sensitive ◽

Exact Nature ◽

Chromatid Cohesion ◽

Lysine Residues

Cohesin is a DNA-associated protein complex that forms a tripartite ring controlling sister chromatid cohesion, chromosome segregation and organization, DNA replication, and gene expression. Sister chromatid cohesion is established by the protein acetyltransferase Eco1, which acetylates two conserved lysine residues on the cohesin subunit Smc3 and thereby ensures correct chromatid separation in yeast (Saccharomyces cerevisiae) and other eukaryotes. However, the consequence of Eco1-catalyzed cohesin acetylation is unknown, and the exact nature of the cohesive state of chromatids remains controversial. Here, we show that self-interactions of the cohesin subunits Scc1/Rad21 and Scc3 occur in a DNA replication–coupled manner in both yeast and human cells. Using cross-linking MS-based and in vivo disulfide cross-linking analyses of purified cohesin, we show that a subpopulation of cohesin may exist as dimers. Importantly, upon temperature-sensitive and auxin-induced degron-mediated Eco1 depletion, the cohesin-cohesin interactions became significantly compromised, whereas deleting either the deacetylase Hos1 or the Eco1 antagonist Wpl1/Rad61 increased cohesin dimer levels by ∼20%. These results indicate that cohesin dimerizes in the S phase and monomerizes in mitosis, processes that are controlled by Eco1, Wpl1, and Hos1 in the sister chromatid cohesion-dissolution cycle. These findings suggest that cohesin dimerization is controlled by the cohesion cycle and support the notion that a double-ring cohesin model operates in sister chromatid cohesion.

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Checkpoint-Dependent and -Independent Roles of Swi3 in Replication Fork Recovery and Sister Chromatid Cohesion in Fission Yeast

PLoS ONE ◽

10.1371/journal.pone.0013379 ◽

2010 ◽

Vol 5 (10) ◽

pp. e13379 ◽

Cited By ~ 13

Author(s):

Jordan B. Rapp ◽

Chiaki Noguchi ◽

Mukund M. Das ◽

Lisa K. Wong ◽

Alison B. Ansbach ◽

...

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Replication Fork ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion

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RFCCtf18 and the Swi1-Swi3 Complex Function in Separate and Redundant Pathways Required for the Stabilization of Replication Forks to Facilitate Sister Chromatid Cohesion in Schizosaccharomyces pombe

Molecular Biology of the Cell ◽

10.1091/mbc.e07-06-0618 ◽

2008 ◽

Vol 19 (2) ◽

pp. 595-607 ◽

Cited By ~ 47

Author(s):

Alison B. Ansbach ◽

Chiaki Noguchi ◽

Ian W. Klansek ◽

Mike Heidlebaugh ◽

Toru M. Nakamura ◽

...

Keyword(s):

Stress Response ◽

Fission Yeast ◽

Sister Chromatid ◽

Replication Fork ◽

Complex Function ◽

Dna Helicase ◽

Sister Chromatid Cohesion ◽

S Phase ◽

Chromatid Cohesion ◽

Factor C

Sister chromatid cohesion is established during S phase near the replication fork. However, how DNA replication is coordinated with chromosomal cohesion pathway is largely unknown. Here, we report studies of fission yeast Ctf18, a subunit of the RFCCtf18 replication factor C complex, and Chl1, a putative DNA helicase. We show that RFCCtf18 is essential in the absence of the Swi1–Swi3 replication fork protection complex required for the S phase stress response. Loss of Ctf18 leads to an increased sensitivity to S phase stressing agents, a decreased level of Cds1 kinase activity, and accumulation of DNA damage during S phase. Ctf18 associates with chromatin during S phase, and it is required for the proper resumption of replication after fork arrest. We also show that chl1Δ is synthetically lethal with ctf18Δ and that a dosage increase of chl1+ rescues sensitivities of swi1Δ to S phase stressing agents, indicating that Chl1 is involved in the S phase stress response. Finally, we demonstrate that inactivation of Ctf18, Chl1, or Swi1-Swi3 leads to defective centromere cohesion, suggesting the role of these proteins in chromosome segregation. We propose that RFCCtf18 and the Swi1–Swi3 complex function in separate and redundant pathways essential for replication fork stabilization to facilitate sister chromatid cohesion in fission yeast.

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Meiotic Chromosome Dynamics Dependent Upon the rec8+, rec10+ and rec11+ Genes of the Fission Yeast Schizosaccharomyces pombe

Genetics ◽

10.1093/genetics/153.1.57 ◽

1999 ◽

Vol 153 (1) ◽

pp. 57-68 ◽

Cited By ~ 1

Author(s):

Michelle D Krawchuk ◽

Linda C DeVeaux ◽

Wayne P Wahls

Keyword(s):

Schizosaccharomyces Pombe ◽

Fission Yeast ◽

Sister Chromatid ◽

Meiotic Chromosome ◽

Sister Chromatid Cohesion ◽

High Rate ◽

Synergistic Interactions ◽

Chromatid Cohesion ◽

Central Regions ◽

Meiosis I

Abstract During meiosis homologous chromosomes replicate once, pair, experience recombination, and undergo two rounds of segregation to produce haploid meiotic products. The rec8+, rec10+, and rec11+ genes of the fission yeast Schizosaccharomyces pombe exhibit similar specificities for meiotic recombination and rec8+ is required for sister chromatid cohesion and homolog pairing. We applied cytological and genetic approaches to identify potential genetic interactions and to gauge the fidelity of meiotic chromosome segregation in the mutants. The rec8+ gene was epistatic to rec10+ and to rec11+, but there was no clear epistatic relationship between rec10+ and rec11+. Reciprocal (crossover) recombination in the central regions of all three chromosomes was compromised in the rec mutants, but recombination near the telomeres was nearly normal. Each of the mutants also exhibited a high rate of aberrant segregation for all three chromosomes. The rec8 mutations affected mainly meiosis I segregation. Remarkably, the rec10 and rec11 mutations, which compromised recombination during meiosis I, affected mainly meiosis II segregation. We propose that these genes encode regulators or components of a “meiotic chromatid cohesion” pathway involved in establishing, maintaining, and appropriately releasing meiotic interactions between chromosomes. A model of synergistic interactions between sister chromatid cohesion and crossover position suggests how crossovers and cohesion help ensure the proper segregation of chromosomes in each of the meiotic divisions.

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