SMC Complexes Are Guarded by the SUMO Protease Ulp2 Against SUMO-Chain-Mediated Turnover

ABSTRACTStructural maintenance of chromosomes (SMC) complexes, cohesin, condensin and Smc5/6, are essential for viability and participate in multiple processes, including sister chromatid cohesion, chromosome condensation, and DNA repair. Here we show that SUMO chains target all three SMC complexes and are antagonized by the SUMO protease Ulp2 to prevent their turnover. We uncover that the essential role of the cohesin-associated subunit Pds5 is to counteract SUMO chains jointly with Ulp2. Importantly, fusion of Ulp2 to kleisin Scc1 supports viability of PDS5 null cells and protects cohesin from proteasomal degradation mediated by the SUMO-targeted ubiquitin ligase Slx5/Slx8. The lethality of PDS5 deleted cells can also be bypassed by simultaneous loss of the PCNA unloader, Elg1, and the cohesin releaser, Wpl1, but only when Ulp2 is functional. Condensin and Smc5/6 complex are similarly guarded by Ulp2 against unscheduled SUMO-chain assembly, which we propose to time the availability of SMC complexes on chromatin.

Sister chromatid cohesion and genome stability in vertebrate cells

Biochemical Society Transactions ◽

10.1042/bst0310263 ◽

2003 ◽

Vol 31 (1) ◽

pp. 263-265 ◽

Cited By ~ 20

Author(s):

C. Morrison ◽

P. Vagnarelli ◽

E. Sonoda ◽

S. Takeda ◽

W.C. Earnshaw

Keyword(s):

Dna Repair ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Genome Stability ◽

Spindle Poles ◽

Chromatid Cohesion ◽

Chromosomal Passenger ◽

Passenger Protein

For successful eukaryotic mitosis, sister chromatid pairs remain linked after replication until their kinetochores have been attached to opposite spindle poles by microtubules. This linkage is broken at the metaphase–anaphase transition and the sisters separate. In budding yeast, this sister chromatid cohesion requires a multi-protein complex called cohesin. A key component of cohesin is Scc1/Mcd1 (Rad21 in fission yeast). Disruption of the chicken orthologue of Scc1 by gene targeting in DT40 cells causes premature sister chromatid separation. Cohesion between sister chromatids is likely to provide a substrate for post-replicative DNA repair by homologous recombination. In keeping with this role of cohesion, Scc1 mutants also show defects in the repair of spontaneous and induced DNA damage. Scc1-deficient cells frequently fail to complete metaphase chromosome alignment and show chromosome segregation defects, suggesting aberrant kinetochore function. Consistent with this, the chromosomal passenger protein, INCENP (inner centromere protein) fails to localize to centromeres. Survivin, another passenger protein and one which interacts with INCENP, also fails to localize to centromeres in Scc1-deficient cells. These results show that cohesin maintains genomic stability by ensuring appropriate DNA repair and equal chromosome segregation at mitosis.

Conserved roles of chromatin remodellers in cohesin loading onto chromatin

Current Genetics ◽

10.1007/s00294-020-01075-x ◽

2020 ◽

Vol 66 (5) ◽

pp. 951-956

Author(s):

Sofía Muñoz ◽

Francesca Passarelli ◽

Frank Uhlmann

Keyword(s):

Sister Chromatid ◽

Domain Architecture ◽

Chromatin Domain ◽

Dual Function ◽

Chromatin Remodelers ◽

Chromatid Cohesion ◽

Highly Expressed Genes ◽

Abstract Cohesin is a conserved, ring-shaped protein complex that topologically entraps DNA. This ability makes this member of the structural maintenance of chromosomes (SMC) complex family a central hub of chromosome dynamics regulation. Besides its essential role in sister chromatid cohesion, cohesin shapes the interphase chromatin domain architecture and plays important roles in transcriptional regulation and DNA repair. Cohesin is loaded onto chromosomes at centromeres, at the promoters of highly expressed genes, as well as at DNA replication forks and sites of DNA damage. However, the features that determine these binding sites are still incompletely understood. We recently described a role of the budding yeast RSC chromatin remodeler in cohesin loading onto chromosomes. RSC has a dual function, both as a physical chromatin receptor of the Scc2/Scc4 cohesin loader complex, as well as by providing a nucleosome-free template for cohesin loading. Here, we show that the role of RSC in sister chromatid cohesion is conserved in fission yeast. We discuss what is known about the broader conservation of the contribution of chromatin remodelers to cohesin loading onto chromatin.

Cohesin dependent compaction of mitotic chromosomes

10.1101/094946 ◽

2016 ◽

Cited By ~ 2

Author(s):

Stephanie A Schalbetter ◽

Anton Goloborodko ◽

Geoffrey Fudenberg ◽

Jon M Belton ◽

Catrina Miles ◽

...

Keyword(s):

Sister Chromatid ◽

Mitotic Chromosome ◽

Protein Complexes ◽

Budding Yeast ◽

Mitotic Chromosomes ◽

Chromosome Conformation ◽

Sister Chromatids ◽

Chromatid Cohesion ◽

Structural Maintenance of Chromosomes (SMC) protein complexes are key determinants of chromosome conformation. Using Hi-C and polymer modelling, we study how cohesin and condensin, two deeply-conserved SMC complexes, organize chromosomes in budding yeast. The canonical role of cohesins is to co-align sister chromatids whilst condensins generally compact mitotic chromosomes. We find strikingly different roles in budding yeast mitosis. First, cohesin is responsible for compacting mitotic chromosomes arms, independent of and in addition to its role in sister-chromatid cohesion. Cohesin dependent mitotic chromosome compaction can be fully accounted for through cis-looping of chromatin by loop extrusion. Second, condensin is dispensable for compaction along chromosomal arms and instead plays a specialized role, structuring rDNA and peri-centromeric regions. Our results argue that the conserved mechanism of SMC complexes is to form chromatin loops and that SMC-dependent looping is readily deployed in a range of contexts to functionally organize chromosomes.

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 ◽

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.

Essential role of Tip60-dependent recruitment of ribonucleotide reductase at DNA damage sites in DNA repair during G1 phase

Genes & Development ◽

10.1101/gad.1863810 ◽

2010 ◽

Vol 24 (4) ◽

pp. 333-338 ◽

Cited By ~ 88

Author(s):

H. Niida ◽

Y. Katsuno ◽

M. Sengoku ◽

M. Shimada ◽

M. Yukawa ◽

...

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Ribonucleotide Reductase ◽

Essential Role ◽

G1 Phase

Organization of Chromosomal DNA by SMC Complexes

Annual Review of Genetics ◽

10.1146/annurev-genet-112618-043633 ◽

2019 ◽

Vol 53 (1) ◽

pp. 445-482 ◽

Cited By ~ 44

Author(s):

Stanislau Yatskevich ◽

James Rhodes ◽

Kim Nasmyth

Keyword(s):

Chromosome Segregation ◽

Sister Chromatid ◽

Mitotic Chromosome ◽

Chromosomal Dna ◽

Chromosome Architecture ◽

Chromatid Cohesion ◽

Dna Translocases ◽

Structural maintenance of chromosomes (SMC) complexes are key organizers of chromosome architecture in all kingdoms of life. Despite seemingly divergent functions, such as chromosome segregation, chromosome maintenance, sister chromatid cohesion, and mitotic chromosome compaction, it appears that these complexes function via highly conserved mechanisms and that they represent a novel class of DNA translocases.

Essential Role of the E3 Ubiquitin Ligase Cbl-b in T Cell Anergy Induction

Immunity ◽

10.1016/j.immuni.2004.07.013 ◽

2004 ◽

Vol 21 (2) ◽

pp. 167-177 ◽

Cited By ~ 244

Author(s):

Myung-Shin Jeon ◽

Alex Atfield ◽

K. Venuprasad ◽

Connie Krawczyk ◽

Renu Sarao ◽

...

Keyword(s):

T Cell ◽

Ubiquitin Ligase ◽

Essential Role ◽

E3 Ubiquitin Ligase ◽

T Cell Anergy ◽

Cell Anergy

The Role of Cdc55 in the Spindle Checkpoint Is through Regulation of Mitotic Exit in Saccharomyces cerevisiae

Molecular Biology of the Cell ◽

10.1091/mbc.e05-04-0336 ◽

2006 ◽

Vol 17 (2) ◽

pp. 658-666 ◽

Cited By ~ 43

Author(s):

Christopher M. Yellman ◽

Daniel J. Burke

Keyword(s):

Sister Chromatid ◽

Spindle Checkpoint ◽

Mitotic Exit ◽

Negative Regulator ◽

Regulatory Subunit ◽

Checkpoint Activation ◽

Chromatid Cohesion ◽

Phosphatase 2A

Cdc55, a B-type regulatory subunit of protein phosphatase 2A, has been implicated in mitotic spindle checkpoint activity and maintenance of sister chromatid cohesion during metaphase. The spindle checkpoint is composed of two independent pathways, one leading to inhibition of the metaphase-to-anaphase transition by checkpoint proteins, including Mad2, and the other to inhibition of mitotic exit by Bub2. We show that Cdc55 is a negative regulator of mitotic exit. A cdc55 mutant, like a bub2 mutant, prematurely releases Cdc14 phosphatase from the nucleolus during spindle checkpoint activation, and premature exit from mitosis indirectly leads to loss of sister chromatid cohesion and inviability in nocodazole. The role of Cdc55 is separable from Bub2 and inhibits release of Cdc14 through a mechanism independent of the known negative regulators of mitotic exit. Epistasis experiments indicate Cdc55 acts either downstream or independent of the mitotic exit network kinase Cdc15. Interestingly, the B-type cyclin Clb2 is partially stable during premature activation of mitotic exit in a cdc55 mutant, indicating mitotic exit is incomplete.