Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA

Xu, Xingya; Kanai, Ryuzi; Nakazawa, Norihiko; Wang, Li; Toyoshima, Chikashi; Yanagida, Mitsuhiro

doi:10.1073/pnas.1803564115

Cited by 20 publications

(59 citation statements)

References 71 publications

(123 reference statements)

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“…Embrace models propose that both sisters are contained within the same cohesin ring, while handcuff models suggest sister chromatids are enclosed in separate rings that are linked together. Recently a new model of chromatin binding by cohesin was reported, termed “hold-and-release” ( Xu et al , 2018 ). The hold-and-release model proposes that DNA is sandwiched by arched coiled-coils of SMC components rather than entrapped within a ring.…”

Section: Discussionmentioning

confidence: 99%

Multiple determinants and consequences of cohesion fatigue in mammalian cells

Sapkota

Wasiak

Daum

et al. 2018

MBoC

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Cells delayed in metaphase with intact mitotic spindles undergo cohesion fatigue, where sister chromatids separate asynchronously, while cells remain in mitosis. Cohesion fatigue requires release of sister chromatid cohesion. However, the pathways that breach sister chromatid cohesion during cohesion fatigue remain unknown. Using moderate-salt buffers to remove loosely bound chromatin cohesin, we show that “cohesive” cohesin is not released during chromatid separation during cohesion fatigue. Using a regulated protein heterodimerization system to lock different cohesin ring interfaces at specific times in mitosis, we show that the Wapl-mediated pathway of cohesin release is not required for cohesion fatigue. By manipulating microtubule stability and cohesin complex integrity in cell lines with varying sensitivity to cohesion fatigue, we show that rates of cohesion fatigue reflect a dynamic balance between spindle pulling forces and resistance to separation by interchromatid cohesion. Finally, while massive separation of chromatids in cohesion fatigue likely produces inviable cell progeny, we find that short metaphase delays, leading to partial chromatid separation, predispose cells to chromosome missegregation. Thus, complete separation of one or a few chromosomes and/or partial separation of sister chromatids may be an unrecognized but common source of chromosome instability that perpetuates the evolution of malignant cells in cancer.

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Section: Discussionmentioning

confidence: 99%

Multiple determinants and consequences of cohesion fatigue in mammalian cells

Sapkota

Wasiak

Daum

et al. 2018

MBoC

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“…The SA cohesin subunit interacts with Rad21. The SMC subunit coiled-coil arms are flexible and can interact with each other forming a rodlike structure and can also fold so that the hinge contacts the head domains [11][12][13][14][15].…”

Section: Cohesin Structure and Chromosome Bindingmentioning

confidence: 99%

The Many Roles of Cohesin in Drosophila Gene Transcription

Dorsett

2019

Trends in Genetics

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The cohesin protein complex mediates sister chromatid cohesion to ensure accurate chromosome segregation, and also influences gene transcription in higher eukaryotes. Modest deficits in cohesin function that do not alter chromosome segregation cause significant birth defects. The mechanisms by which cohesin participates in gene regulation have been studied in Drosophila, revealing that it is involved in gene activation by transcriptional enhancers and epigenetic gene silencing by Polycomb group proteins. Recent studies reveal that early DNA replication origins are important for determining which genes associate with cohesin and suggest that cohesin at replication origins is important for establishing both sister chromatid cohesion and enhancerpromoter communication.

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“…8B). Even more striking, the very same mutations were recovered as intragenic suppressors of the ts allele mis4-G1326E [56] suggesting they enhance Mis4 activity. This region is enriched for residues mutated in Cornelia de Lange syndrome and Kikuchi et al have shown that many of these mutations specifically disrupt the Scc2-Scc1 interaction ( [19] and Fig.…”

Section: Rad21 Phosphorylation Status May Modulate the Activity Of Thmentioning

confidence: 88%

The CDK Pef1 and Protein Phosphatase 4 oppose each other for regulating cohesin binding to fission yeast chromosomes

Birot

Tormos-Pérez

Vaur

et al. 2019

Preprint

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Cohesin has essential roles in chromosome structure, segregation and repair. Cohesin binding to chromosomes is catalyzed by the cohesin loader, Mis4 in fission yeast. How cells fine tune cohesin deposition is largely unknown. Here we provide evidence that Mis4 activity is regulated by phosphorylation of its cohesin substrate. A genetic screen for negative regulators of Mis4 yielded a CDK called Pef1, whose closest human homologue is CDK5. Inhibition of Pef1 kinase activity rescued cohesin loader deficiencies. In an otherwise wildtype background, Pef1 ablation stimulated cohesin binding to its regular sites along chromosomes while ablating Protein Phosphatase 4 had the opposite effect. Pef1 and PP4 control the phosphorylation state of the cohesin kleisin Rad21. The CDK phosphorylates Rad21 on Threonine 262. Pef1 ablation, non phosphorylatable Rad21-T262 or mutations within a Rad21 binding domain of Mis4 alleviated the effect of PP4 deficiency. Such a CDK/PP4 based regulation of cohesin loader activity could provide an efficient mechanism for translating cellular cues into a fast and accurate cohesin response.3 DNA capture may be achieved by modulating the catalytic activity of the loader and concerted transcriptional responses may involve a local and temporal control of loading and unloading activities. How cells orchestrate cohesin functions is largely unknown. Intriguingly, the kleisin subunit of cohesin is targeted by multiple phosphorylation events. In fission yeast, Rad21 shows multiple phospho-isoforms whose relative abundance fluctuates along the cell cycle [39]. Our recent work showed that Protein Phosphatase 4 (PP4) controls the phosphorylation status of Rad21 and modulates Wpl1 activity [40], leading to the idea that cohesin functions could be spatially and temporally fine-tuned by altering the balance between kinase and phosphatase activities.Here we report on the control of cohesin deposition by the opposite activities of the Pef1 CDK and PP4. Pef1 was first described as a PSTAIRE-related protein in fission yeast [41]. The CDK has three known cyclin partners called Pas1, Psl1 and Clg1 and was reported to facilitate the G1 to S phase transition and to regulate life span [42; 43]. Its closest human homolog, CDK5, is involved in a myriad of cellular functions and pathologies, from neurodegenerative diseases to multiple solid and hematological cancers [44]. We identified pef1 in a genetic screen for mutants able to rescue the cohesin loader mutant mis4-367. Pef1 ablation or inhibition of its kinase activity increased cohesin deposition and rescued sister-chromatid cohesion defects of the mis4 mutant. In otherwise wild-type cells Pef1 ablation increased the binding of both cohesin and its loader to their regular sites along chromosomes. Genetic analyses indicated that Pef1 acts through the phosphorylation of multiple targets. We identified one of these within the kleisin Rad21. Specifically, the Pef1/Psl1 complex phosphorylates Rad21 on T262 and preventing this phosphorylation event recapitulates in part th...

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Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA

Cited by 20 publications

References 71 publications

Multiple determinants and consequences of cohesion fatigue in mammalian cells

Multiple determinants and consequences of cohesion fatigue in mammalian cells

The Many Roles of Cohesin in Drosophila Gene Transcription

The CDK Pef1 and Protein Phosphatase 4 oppose each other for regulating cohesin binding to fission yeast chromosomes

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