Polo kinase Cdc5 associates with centromeres to facilitate the removal of centromeric cohesin during mitosis

Sister chromatid cohesion is essential for tension-sensing mechanisms that monitor bipolar attachment of replicated chromatids in metaphase. Cohesion is mediated by the association of cohesins along the length of sister chromatid arms. In contrast, centromeric cohesin generates intrastrand cohesion and sister centromeres, while highly cohesin enriched, are separated by >800 nm at metaphase in yeast. Removal of cohesin is necessary for sister chromatid separation during anaphase, and this is regulated by evolutionarily conserved polo-like kinase (Cdc5 in yeast, Plk1 in humans). Here we address how high levels of cohesins at centromeric chromatin are removed. Cdc5 associates with centromeric chromatin and cohesin-associated regions. Maximum enrichment of Cdc5 in centromeric chromatin occurs during the metaphase-to-anaphase transition and coincides with the removal of chromosome-associated cohesin. Cdc5 interacts with cohesin in vivo, and cohesin is required for association of Cdc5 at centromeric chromatin. Cohesin removal from centromeric chromatin requires Cdc5 but removal at distal chromosomal arm sites does not. Our results define a novel role for Cdc5 in regulating removal of centromeric cohesins and faithful chromosome segregation.

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A quantitative analysis of cohesin decay in mitotic fidelity

The Journal of Cell Biology ◽

10.1083/jcb.201801111 ◽

2018 ◽

Vol 217 (10) ◽

pp. 3343-3353 ◽

Cited By ~ 6

Author(s):

Sara Carvalhal ◽

Alexandra Tavares ◽

Mariana B. Santos ◽

Mihailo Mirkovic ◽

Raquel A. Oliveira

Keyword(s):

Quantitative Analysis ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Mitotic Spindle ◽

Sister Chromatid Cohesion ◽

Force Balance ◽

Chromosome Organization ◽

Centromeric Chromatin ◽

Chromatid Cohesion ◽

Chromosome Alignment

Sister chromatid cohesion mediated by cohesin is essential for mitotic fidelity. It counteracts spindle forces to prevent premature chromatid individualization and random genome segregation. However, it is unclear what effects a partial decline of cohesin may have on chromosome organization. In this study, we provide a quantitative analysis of cohesin decay by inducing acute removal of defined amounts of cohesin from metaphase-arrested chromosomes. We demonstrate that sister chromatid cohesion is very resistant to cohesin loss as chromatid disjunction is only observed when chromosomes lose >80% of bound cohesin. Removal close to this threshold leads to chromosomes that are still cohered but display compromised chromosome alignment and unstable spindle attachments. Partial cohesin decay leads to increased duration of mitosis and susceptibility to errors in chromosome segregation. We propose that high cohesin density ensures centromeric chromatin rigidity necessary to maintain a force balance with the mitotic spindle. Partial cohesin loss may lead to chromosome segregation errors even when sister chromatid cohesion is fulfilled.

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Multiple Subunits of the Caenorhabditis elegans Anaphase-Promoting Complex Are Required for Chromosome Segregation During Meiosis I

Genetics ◽

10.1093/genetics/160.2.805 ◽

2002 ◽

Vol 160 (2) ◽

pp. 805-813 ◽

Cited By ~ 3

Author(s):

Edward S Davis ◽

Lucia Wille ◽

Barry A Chestnut ◽

Penny L Sadler ◽

Diane C Shakes ◽

...

Keyword(s):

Rna Interference ◽

Caenorhabditis Elegans ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Anaphase Promoting Complex ◽

Genetic Screens ◽

Chromatid Cohesion ◽

Interference Studies ◽

Meiosis I

Abstract Two genes, originally identified in genetic screens for Caenorhabditis elegans mutants that arrest in metaphase of meiosis I, prove to encode subunits of the anaphase-promoting complex or cyclosome (APC/C). RNA interference studies reveal that these and other APC/C subunits are essential for the segregation of chromosomal homologs during meiosis I. Further, chromosome segregation during meiosis I requires APC/C functions in addition to the release of sister chromatid cohesion.

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Double or nothing: a Drosophila mutation affecting meiotic chromosome segregation in both females and males.

Genetics ◽

10.1093/genetics/136.3.953 ◽

1994 ◽

Vol 136 (3) ◽

pp. 953-964 ◽

Cited By ~ 2

Author(s):

D P Moore ◽

W Y Miyazaki ◽

J E Tomkiel ◽

T L Orr-Weaver

Keyword(s):

Cell Death ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Meiotic Chromosome ◽

Sister Chromatid Cohesion ◽

Aberrant Chromosome ◽

Chromatid Cohesion ◽

Meiotic Nondisjunction ◽

Lethal Allele ◽

Meiosis I

Abstract We describe a Drosophila mutation, Double or nothing (Dub), that causes meiotic nondisjunction in a conditional, dominant manner. Previously isolated mutations in Drosophila specifically affect meiosis either in females or males, with the exception of the mei-S332 and ord genes which are required for proper sister-chromatid cohesion. Dub is unusual in that it causes aberrant chromosome segregation almost exclusively in meiosis I in both sexes. In Dub mutant females both nonexchange and exchange chromosomes undergo nondisjunction, but the effect of Dub on nonexchange chromosomes is more pronounced. Dub reduces recombination levels slightly. Multiple nondisjoined chromosomes frequently cosegregate to the same pole. Dub results in nondisjunction of all chromosomes in meiosis I of males, although the levels are lower than in females. When homozygous, Dub is a conditional lethal allele and exhibits phenotypes consistent with cell death.

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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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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 ◽

Sister Chromatid Cohesion ◽

Chromosomal Dna ◽

Chromosome Architecture ◽

Chromatid Cohesion ◽

Dna Translocases ◽

Structural Maintenance Of Chromosomes

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.

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The acetyltransferase activity of San stabilizes the mitotic cohesin at the centromeres in a shugoshin-independent manner

The Journal of Cell Biology ◽

10.1083/jcb.200701043 ◽

2007 ◽

Vol 177 (4) ◽

pp. 587-597 ◽

Cited By ~ 55

Author(s):

Fajian Hou ◽

Chih-Wen Chu ◽

Xiangduo Kong ◽

Kyoko Yokomori ◽

Hui Zou

Keyword(s):

Cell Cycle ◽

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Independent Manner ◽

Acetyltransferase Activity ◽

Chromatid Cohesion ◽

Interphase Cells ◽

Gain Access

Proper sister chromatid cohesion is critical for maintaining genetic stability. San is a putative acetyltransferase that is important for sister chromatid cohesion in Drosophila melanogaster, but not in budding yeast. We showed that San is critical for sister chromatid cohesion in HeLa cells, suggesting that this mechanism may be conserved in metazoans. Furthermore, although a small fraction of San interacts with the NatA complex, San appears to mediate cohesion independently. San exhibits acetyltransferase activity in vitro, and its activity is required for sister chromatid cohesion in vivo. In the absence of San, Sgo1 localizes correctly throughout the cell cycle. However, cohesin is no longer detected at the mitotic centromeres. Furthermore, San localizes to the cytoplasm in interphase cells; thus, it may not gain access to chromosomes until mitosis. Moreover, in San-depleted cells, further depletion of Plk1 rescues the cohesion along the chromosome arms, but not at the centromeres. Collectively, San may be specifically required for the maintenance of the centromeric cohesion in mitosis.

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NudCL2 is an Hsp90 cochaperone to regulate sister chromatid cohesion by stabilizing cohesin subunits

10.1101/358911 ◽

2018 ◽

Author(s):

Yuehong Yang ◽

Wei Wang ◽

Min Li ◽

Wen Zhang ◽

Yuliang Huang ◽

...

Keyword(s):

Atpase Activity ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Mammalian Cells ◽

Ectopic Expression ◽

Sister Chromatid Cohesion ◽

Chaperone Function ◽

Chromatid Cohesion ◽

Protein Instability ◽

The Stability

AbstractSister chromatid cohesion plays a key role in ensuring precise chromosome segregation during mitosis, which is mediated by the multisubunit complex cohesin. However, the molecular regulation of cohesin subunits stability remains unclear. Here, we show that NudCL2 (NudC-like protein 2) is essential for the stability of cohesin subunits by regulating Hsp90 ATPase activity in mammalian cells. Depletion of NudCL2 induces mitotic defects and premature sister chromatid separation and destabilizes cohesin subunits that interact with NudCL2. Similar defects are also observed upon inhibition of Hsp90 ATPase activity. Interestingly, ectopic expression of Hsp90 efficiently rescues the protein instability and functional deficiency of cohesin induced by NudCL2 depletion, but not vice versa. Moreover, NudCL2 not only binds to Hsp90, but also significantly modulates Hsp90 ATPase activity and promotes the chaperone function of Hsp90. Taken together, these data suggest that NudCL2 is a previously undescribed Hsp90 cochaperone to modulate sister chromatid cohesion by stabilizing cohesin subunits, providing a hitherto unrecognized mechanism that is crucial for faithful chromosome segregation during mitosis.

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Defects in Components of the Proteasome Enhance Transcriptional Silencing at Fission Yeast Centromeres and Impair Chromosome Segregation

Molecular and Cellular Biology ◽

10.1128/mcb.19.7.5155 ◽

1999 ◽

Vol 19 (7) ◽

pp. 5155-5165 ◽

Cited By ~ 12

Author(s):

Jean-Paul Javerzat ◽

Gordon McGurk ◽

Gwen Cranston ◽

Christian Barreau ◽

Pascal Bernard ◽

...

Keyword(s):

Fission Yeast ◽

Sister Chromatid ◽

Position Effect ◽

Transcriptional Silencing ◽

Position Effect Variegation ◽

Centromeric Chromatin ◽

Centromere Function ◽

Chromatid Cohesion ◽

Sister Chromatid Separation ◽

Elevated Rate

ABSTRACT Fission yeast centromeres are transcriptionally silent and form a heterochromatin-like structure essential for normal centromere function; this appears analogous to heterochromatin and position effect variegation in other eukaryotes. Conditional mutations in three genes designated cep (centromere enhancer of position effect) were found to enhance transcriptional silencing within centromeres. Cloning of the cep1 + andcep2 + genes by functional complementation revealed that they are identical to the previously described genespad1 + and mts2 +, respectively, which both encode subunits of the proteasome 19S cap. Like Mts2 and Mts4, epitope-tagged Cep1/Pad1 localizes to or near the nuclear envelope throughout the cell cycle. The cep mutants display a range of phenotypes depending on the temperature. Silencing within the central domain of centromeres is increased at 36°C. This suggests that the proteasome is involved in regulating silencing and thus centromeric chromatin architecture, possibly by lowering the level of some chromatin-associated protein by ubiquitin-dependent degradation. This is the first report of defective proteasome function affecting heterochromatin-mediated transcriptional silencing. At 36 and 32°C, the cep mutants lose chromosomes at an elevated rate, and at 18°C, the mutants are cryosensitive for growth. Cytological analysis at 18°C revealed a defect in sister chromatid separation while other mitotic events occurred normally, indicating that cep mutations might interfere specifically with the degradation of inhibitor(s) of sister chromatid separation. These observations suggest that 19S subunits confer a level of substrate specificity on the proteasome and raise the possibility of a link between components involved in centromere architecture and sister chromatid cohesion.

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Acetylation of the SUN protein Mps3 by Eco1 regulates its function in nuclear organization

Molecular Biology of the Cell ◽

10.1091/mbc.e11-07-0600 ◽

2012 ◽

Vol 23 (13) ◽

pp. 2546-2559 ◽

Cited By ~ 15

Author(s):

Suman Ghosh ◽

Jennifer M. Gardner ◽

Christine J. Smoyer ◽

Jennifer M. Friederichs ◽

Jay R. Unruh ◽

...

Keyword(s):

Nuclear Membrane ◽

Sister Chromatid ◽

Transmembrane Domain ◽

Nuclear Organization ◽

Spindle Pole Body ◽

Spindle Pole ◽

Sister Chromatid Cohesion ◽

Chromatid Cohesion

The Saccharomyces cerevisiae SUN-domain protein Mps3 is required for duplication of the yeast centrosome-equivalent organelle, the spindle pole body (SPB), and it is involved in multiple aspects of nuclear organization, including telomere tethering and gene silencing at the nuclear membrane, establishment of sister chromatid cohesion, and repair of certain types of persistent DNA double-stranded breaks. How these diverse SUN protein functions are regulated is unknown. Here we show that the Mps3 N-terminus is a substrate for the acetyltransferase Eco1/Ctf7 in vitro and in vivo and map the sites of acetylation to three lysine residues adjacent to the Mps3 transmembrane domain. Mutation of these residues shows that acetylation is not essential for growth, SPB duplication, or distribution in the nuclear membrane. However, analysis of nonacetylatable mps3 mutants shows that this modification is required for accurate sister chromatid cohesion and for chromosome recruitment to the nuclear membrane. Acetylation of Mps3 by Eco1 is one of the few regulatory mechanisms known to control nuclear organization.

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Pericentromeric Sister Chromatid Cohesion Promotes Kinetochore Biorientation

Molecular Biology of the Cell ◽

10.1091/mbc.e09-04-0330 ◽

2009 ◽

Vol 20 (17) ◽

pp. 3818-3827 ◽

Cited By ~ 65

Author(s):

Tessie M. Ng ◽

William G. Waples ◽

Brigitte D. Lavoie ◽

Sue Biggins

Keyword(s):

Protein Kinase ◽

Genetic Analysis ◽

Chromosome Segregation ◽

Sister Chromatid ◽

Budding Yeast ◽

Sister Chromatid Cohesion ◽

Specific Role ◽

Aurora B ◽

Chromatid Cohesion ◽

Yeast Genes

Accurate chromosome segregation depends on sister kinetochores making bioriented attachments to microtubules from opposite poles. An essential regulator of biorientation is the Ipl1/Aurora B protein kinase that destabilizes improper microtubule–kinetochore attachments. To identify additional biorientation pathways, we performed a systematic genetic analysis between the ipl1-321 allele and all nonessential budding yeast genes. One of the mutants, mcm21Δ, precociously separates pericentromeres and this is associated with a defect in the binding of the Scc2 cohesin-loading factor at the centromere. Strikingly, Mcm21 becomes essential for biorientation when Ipl1 function is reduced, and this appears to be related to its role in pericentromeric cohesion. When pericentromeres are artificially tethered, Mcm21 is no longer needed for biorientation despite decreased Ipl1 activity. Taken together, these data reveal a specific role for pericentromeric linkage in ensuring kinetochore biorientation.

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