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

ABSTRACT Sister chromatid cohesion is essential for cell viability. We have isolated a novel temperature-sensitive lethal mutant namedeso1-H17 that displays spindle assembly checkpoint-dependent mitotic delay and abnormal chromosome segregation. At the permissive temperature, the eso1-H17 mutant shows mild sensitivity to UV irradiation and DNA-damaging chemicals. At the nonpermissive temperature, the mutant is arrested in M phase with a viability loss due to a failure to establish sister chromatid cohesion during S phase. The lethal M-phase arrest phenotype, however, is suppressed by inactivation of a spindle checkpoint. Theeso1 + gene is not essential for the onset and progression of DNA replication but has remarkable genetic interactions with those genes regulating the G1-S transition and DNA replication. The N-terminal two-thirds of Eso1p is highly homologous to DNA polymerase η of budding yeast and humans, and the C-terminal one-third is homologous to budding yeast Eco1p (also called Ctf7p), which is required for the establishment of sister chromatid cohesion. Deletion analysis and determination of the mutation site reveal that the function of the Eco1p/Ctf7p-homologous domain is necessary and sufficient for sister chromatid cohesion. On the other hand, deletion of the DNA polymerase η domain in Eso1p increases sensitivity to UV irradiation. These results indicate that Eso1p plays a dual role during DNA replication. The C-terminal region acts to establish sister chromatid cohesion, and the N-terminal region presumably catalyzes translesion DNA synthesis when template DNA contains lesions that block regular DNA replication.

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A Coordinated Temporal Interplay of Nucleosome Reorganization Factor, Sister Chromatin Cohesion Factor, and DNA Polymerase α Facilitates DNA Replication

Molecular and Cellular Biology ◽

10.1128/mcb.24.21.9568-9579.2004 ◽

2004 ◽

Vol 24 (21) ◽

pp. 9568-9579 ◽

Cited By ~ 43

Author(s):

Yanjiao Zhou ◽

Teresa S.-F. Wang

Keyword(s):

Dna Replication ◽

Dna Polymerase ◽

S Phase ◽

Terminal Region ◽

Replication Complex ◽

Dna Polymerase Α ◽

Chromatid Cohesion ◽

Phase Progression

ABSTRACT DNA replication depends critically upon chromatin structure. Little is known about how the replication complex overcomes the nucleosome packages in chromatin during DNA replication. To address this question, we investigate factors that interact in vivo with the principal initiation DNA polymerase, DNA polymerase α (Polα). The catalytic subunit of budding yeast Polα (Pol1p) has been shown to associate in vitro with the Spt16p-Pob3p complex, a component of the nucleosome reorganization system required for both replication and transcription, and with a sister chromatid cohesion factor, Ctf4p. Here, we show that an N-terminal region of Polα (Pol1p) that is evolutionarily conserved among different species interacts with Spt16p-Pob3p and Ctf4p in vivo. A mutation in a glycine residue in this N-terminal region of POL1 compromises the ability of Pol1p to associate with Spt16p and alters the temporal ordered association of Ctf4p with Pol1p. The compromised association between the chromatin-reorganizing factor Spt16p and the initiating DNA polymerase Pol1p delays the Pol1p assembling onto and disassembling from the late-replicating origins and causes a slowdown of S-phase progression. Our results thus suggest that a coordinated temporal and spatial interplay between the conserved N-terminal region of the Polα protein and factors that are involved in reorganization of nucleosomes and promoting establishment of sister chromatin cohesion is required to facilitate S-phase progression.

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Correct spindle elongation at the metaphase/anaphase transition is an APC-dependent event in budding yeast

The Journal of Cell Biology ◽

10.1083/jcb.200104096 ◽

2001 ◽

Vol 155 (5) ◽

pp. 711-718 ◽

Cited By ~ 25

Author(s):

Fedor Severin ◽

Anthony A. Hyman ◽

Simonetta Piatti

Keyword(s):

Cell Cycle ◽

Dna Replication ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Budding Yeast ◽

Sister Chromatid Cohesion ◽

Anaphase Promoting Complex ◽

Sister Chromatids ◽

Chromatid Cohesion ◽

Spindle Elongation

At the metaphase to anaphase transition, chromosome segregation is initiated by the splitting of sister chromatids. Subsequently, spindles elongate, separating the sister chromosomes into two sets. Here, we investigate the cell cycle requirements for spindle elongation in budding yeast using mutants affecting sister chromatid cohesion or DNA replication. We show that separation of sister chromatids is not sufficient for proper spindle integrity during elongation. Rather, successful spindle elongation and stability require both sister chromatid separation and anaphase-promoting complex activation. Spindle integrity during elongation is dependent on proteolysis of the securin Pds1 but not on the activity of the separase Esp1. Our data suggest that stabilization of the elongating spindle at the metaphase to anaphase transition involves Pds1-dependent targets other than Esp1.

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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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S-Phase Cyclin-Dependent Kinases Promote Sister Chromatid Cohesion in Budding Yeast

Molecular and Cellular Biology ◽

10.1128/mcb.05323-11 ◽

2011 ◽

Vol 31 (12) ◽

pp. 2470-2483 ◽

Cited By ~ 8

Author(s):

W.- S. Hsu ◽

S. L. Erickson ◽

H.- J. Tsai ◽

C. A. Andrews ◽

A. C. Vas ◽

...

Keyword(s):

Sister Chromatid ◽

Budding Yeast ◽

Sister Chromatid Cohesion ◽

S Phase ◽

Cyclin Dependent Kinases ◽

Chromatid Cohesion

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The acetyltransferase Eco1 elicits cohesin dimerization during S phase

10.1101/2020.02.07.938530 ◽

2020 ◽

Author(s):

Di Shi ◽

Shuaijun Zhao ◽

Mei-Qing Zuo ◽

Jingjing Zhang ◽

Wenya Hou ◽

...

Keyword(s):

Dna Replication ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

S Phase ◽

Repair Gene ◽

Additional Layer ◽

Ring Model ◽

Dna Repair Gene ◽

Chromatid Cohesion

AbstractSister chromatid cohesion is established by Eco1 in S phase. Nevertheless, the exact consequence of Eco1-catalyzed acetylation is unknown, and the cohesive state remains highly controversial. Here we show that self-interactions of cohesin subunits Scc1/Rad21 and Scc3 occur in a DNA replication-coupled manner in both yeast and human. Through cross-linking mass spectrometry and VivosX analysis of purified cohesin, we show that a subpopulation of cohesin may exist as dimers. Importantly, cohesin-cohesin interaction becomes significantly compromised when Eco1 is depleted. On the other hand, deleting either deacetylase Hos1 or Eco1 antagonist Wpl1/Rad61 results in an increase (e.g., from ∼20% to 40%) of cohesin dimers. These findings suggest that cohesin dimerization is controlled by common mechanisms as the cohesion cycle, thus providing an additional layer of regulation for cohesin to execute various functions such as sister chromatid cohesion, DNA repair, gene expression, chromatin looping and high-order organization.Author SummaryCohesin is a ring that tethers sister chromatids since their synthesis during S phase till their separation in anaphase. According to the single-ring model, one ring holds twin sisters. Here we show a conserved cohesin-cohesin interaction from yeast to human. A subpopulation of cohesin is dimerized concomitantly with DNA replication. Cohesin dimerization is dependent on the acetyltransferase Eco1 and counteracted by the anti-establishment factor Wpl1 and deacetylase Hos1. Approximately 20% of cellular cohesin complexes are measured to be dimers, close to the level of Smc3 acetylation by Eco1 in vivo. These findings provide evidence to support the double-ring model in sister chromatid cohesion.

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Pol kappa : A DNA Polymerase Required for Sister Chromatid Cohesion

Science ◽

10.1126/science.289.5480.774 ◽

2000 ◽

Vol 289 (5480) ◽

pp. 774-779 ◽

Cited By ~ 130

Author(s):

Z. Wang

Keyword(s):

Dna Polymerase ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion

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Cul4-Ddb1 ubiquitin ligases facilitate DNA replication-coupled sister chromatid cohesion through regulation of cohesin acetyltransferase Esco2

10.1101/414888 ◽

2018 ◽

Cited By ~ 1

Author(s):

Haitao Sun ◽

Jiaxin Zhang ◽

Jingjing Zhang ◽

Zhen Li ◽

Qinhong Cao ◽

...

Keyword(s):

Dna Replication ◽

Ubiquitin Ligase ◽

Sister Chromatid ◽

Critical Role ◽

E3 Ubiquitin Ligase ◽

Ubiquitin Ligases ◽

Sister Chromatid Cohesion ◽

Vital Role ◽

Chromatid Cohesion ◽

Evolutionarily Conserved

AbstractCohesin acetyltransferases Esco1 and Esco2 play a vital role in establishing sister chromatid cohesion. How Esco1 and Esco2 are controlled to achieve this in a DNA replication-coupled manner remains unclear in higher eukaryotes. Here we show that Cul4-RING ligases (CRL4s) play a critical role in sister chromatid cohesion in human cells. Depletion of Cul4A, Cul4B or Ddb1 subunits substantially reduces normal cohesion efficiency. We also show that Mms22L, a vertebrate ortholog of yeast Mms22, is one of Ddb1 and Cul4-associated factors (DCAFs) involved in cohesion. Several lines of evidence suggest a selective interaction of CRL4s with Esco2, but not Esco1. Depletion of either CRL4s or Esco2 causes a defect in Smc3 acetylation which can be rescued by HDAC8 inhibition. More importantly, both CRL4s and PCNA act as mediators for efficiently stabilizing Esco2 on chromatin and catalyzing Smc3 acetylation. Taken together, we propose an evolutionarily conserved mechanism in which CRL4s and PCNA regulate Esco2-dependent establishment of sister chromatid cohesion.Author summaryWe identified human Mms22L as a substrate specific adaptor of Cul4-Ddb1 E3 ubiquitin ligase. Downregulation of Cul4A, Cul4B or Ddb1 subunit causes reduction of acetylated Smc3, via interaction with Esco2 acetyltransferase, and then impairs sister chromatid cohesion in 293T cells. We found functional complementation between Cul4-Ddb1-Mms22L E3 ligase and Esco2 in Smc3 acetylation and sister chromatid cohesion. Interestingly, both Cul4-Ddb1 E3 ubiquitin ligase and PCNA contribute to Esco2 mediated Smc3 acetylation. To summarise, we demonstrated an evolutionarily conserved mechanism in which Cul4-Ddb1 E3 ubiquitin ligases and PCNA regulate Esco2-dependent establishment of sister chromatid cohesion.

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Fission yeast Pds5 is required for accurate chromosome segregation and for survival after DNA damage or metaphase arrest

Journal of Cell Science ◽

10.1242/jcs.115.3.587 ◽

2002 ◽

Vol 115 (3) ◽

pp. 587-598 ◽

Cited By ~ 3

Author(s):

Shao-Win Wang ◽

Rebecca L. Read ◽

Chris J. Norbury

Keyword(s):

Dna Damage ◽

Fission Yeast ◽

Sister Chromatid ◽

Budding Yeast ◽

Eukaryotic Cell ◽

Sister Chromatid Cohesion ◽

Chromosome Loss ◽

Dependent Manner ◽

Yeast Saccharomyces Cerevisiae ◽

Chromatid Cohesion

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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