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

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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Faculty Opinions recommendation of Establishment of sister chromatid cohesion at the S. cerevisiae replication fork.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1041064.492943 ◽

2006 ◽

Author(s):

Fyodor Urnov

Keyword(s):

Sister Chromatid ◽

Replication Fork ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion

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Interaction of the Warsaw breakage syndrome DNA helicase DDX11 with the replication fork-protection factor Timeless promotes sister chromatid cohesion

PLoS Genetics ◽

10.1371/journal.pgen.1007622 ◽

2018 ◽

Vol 14 (10) ◽

pp. e1007622 ◽

Cited By ~ 13

Author(s):

Giuseppe Cortone ◽

Ge Zheng ◽

Pasquale Pensieri ◽

Viviana Chiappetta ◽

Rosarita Tatè ◽

...

Keyword(s):

Sister Chromatid ◽

Replication Fork ◽

Dna Helicase ◽

Sister Chromatid Cohesion ◽

Protection Factor ◽

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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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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Establishment of Sister Chromatid Cohesion at the S. cerevisiae Replication Fork

Molecular Cell ◽

10.1016/j.molcel.2006.08.018 ◽

2006 ◽

Vol 23 (6) ◽

pp. 787-799 ◽

Cited By ~ 198

Author(s):

Armelle Lengronne ◽

John McIntyre ◽

Yuki Katou ◽

Yutaka Kanoh ◽

Karl-Peter Hopfner ◽

...

Keyword(s):

Sister Chromatid ◽

Replication Fork ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion

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Towards a Structural Mechanism for Sister Chromatid Cohesion Establishment at the Eukaryotic Replication Fork

Biology ◽

10.3390/biology10060466 ◽

2021 ◽

Vol 10 (6) ◽

pp. 466

Author(s):

Sarah S. Henrikus ◽

Alessandro Costa

Keyword(s):

Single Molecule ◽

Sister Chromatid ◽

Replication Fork ◽

Genetic Material ◽

Sister Chromatid Cohesion ◽

Duplex Dna ◽

Structural Mechanism ◽

Chromatin Dynamics ◽

Chromatid Cohesion ◽

Replication Factors

Cohesion between replicated chromosomes is essential for chromatin dynamics and equal segregation of duplicated genetic material. In the G1 phase, the ring-shaped cohesin complex is loaded onto duplex DNA, enriching at replication start sites, or “origins”. During the same phase of the cell cycle, and also at the origin sites, two MCM helicases are loaded as symmetric double hexamers around duplex DNA. During the S phase, and through the action of replication factors, cohesin switches from encircling one parental duplex DNA to topologically enclosing the two duplicated DNA filaments, which are known as sister chromatids. Despite its vital importance, the structural mechanism leading to sister chromatid cohesion establishment at the replication fork is mostly elusive. Here we review the current understanding of the molecular interactions between the replication machinery and cohesin, which support sister chromatid cohesion establishment and cohesin function. In particular, we discuss how cryo-EM is shedding light on the mechanisms of DNA replication and cohesin loading processes. We further expound how frontier cryo-EM approaches, combined with biochemistry and single-molecule fluorescence assays, can lead to understanding the molecular basis of sister chromatid cohesion establishment at the replication fork.

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

Molecular and Cellular Biology ◽

10.1128/mcb.01284-10 ◽

2011 ◽

Vol 31 (8) ◽

pp. 1771-1786 ◽

Cited By ~ 40

Author(s):

A. Feytout ◽

S. Vaur ◽

S. Genier ◽

S. Vazquez ◽

J.-P. Javerzat

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion ◽

Lysine Residues

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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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Saccharomyces cerevisiae CTF18 and CTF4 Are Required for Sister Chromatid Cohesion

Molecular and Cellular Biology ◽

10.1128/mcb.21.9.3144-3158.2001 ◽

2001 ◽

Vol 21 (9) ◽

pp. 3144-3158 ◽

Cited By ~ 218

Author(s):

Joseph S. Hanna ◽

Evgueny S. Kroll ◽

Victoria Lundblad ◽

Forrest A. Spencer

Keyword(s):

Sister Chromatid ◽

Replication Fork ◽

Spindle Assembly Checkpoint ◽

Budding Yeast ◽

Sister Chromatid Cohesion ◽

Nuclear Antigen ◽

Chromatid Cohesion ◽

Core Components ◽

Proliferating Cell

ABSTRACT CTF4 and CTF18 are required for high-fidelity chromosome segregation. Both exhibit genetic and physical ties to replication fork constituents. We find that absence of eitherCTF4 or CTF18 causes sister chromatid cohesion failure and leads to a preanaphase accumulation of cells that depends on the spindle assembly checkpoint. The physical and genetic interactions between CTF4, CTF18, and core components of replication fork complexes observed in this study and others suggest that both gene products act in association with the replication fork to facilitate sister chromatid cohesion. We find that Ctf18p, anRFC1-like protein, directly interacts with Rfc2p, Rfc3p, Rfc4p, and Rfc5p. However, Ctf18p is not a component of biochemically purified proliferating cell nuclear antigen loading RF-C, suggesting the presence of a discrete complex containing Ctf18p, Rfc2p, Rfc3p, Rfc4p, and Rfc5p. Recent identification and characterization of the budding yeast polymerase κ, encoded by TRF4, strongly supports a hypothesis that the DNA replication machinery is required for proper sister chromatid cohesion. Analogous to the polymerase switching role of the bacterial and human RF-C complexes, we propose that budding yeast RF-CCTF18 may be involved in a polymerase switch event that facilities sister chromatid cohesion. The requirement for CTF4 and CTF18 in robust cohesion identifies novel roles for replication accessory proteins in this process.

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