Functions of the centromere and kinetochore in chromosome segregation

Westhorpe, Frederick G.; Straight, Aaron F.

doi:10.1016/j.ceb.2013.02.001

Cited by 113 publications

(90 citation statements)

References 61 publications

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“…However, this tethering does not serve to recruit the Ndc80 complex during kinetochore assembly as cells lacking Cnn1 do not suffer from reduced levels of the Ndc80 complex, nor a reduction in the Mtw1 and Spc105 complexes (Bock et al 2012). In contrast, mammalian CENP-T actively recruits the Ndc80 complex to CENs (Gascoigne et al 2011) and the middle region of CENP-T appears to be flexible and assists in kinetochore stretching when it undergoes tension (Perpelescu and Fukagawa 2011;Suzuki et al 2011;Westhorpe and Straight 2013). Similarly, the preanaphase Cnn1 linkages may allow a proper intrakinetochore stretch required for chromosome biorientation, as shown for CENP-T (Suzuki et al 2014).…”

Section: Discussionmentioning

confidence: 99%

The Mps1 Kinase Modulates the Recruitment and Activity of Cnn1CENP-T at Saccharomyces cerevisiae Kinetochores

et al. 2015

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Kinetochores are conserved protein complexes that bind the replicated chromosomes to the mitotic spindle and then direct their segregation. To better comprehend Saccharomyces cerevisiae kinetochore function, we dissected the phospho-regulated dynamic interaction between conserved kinetochore protein Cnn1 CENP-T , the centromere region, and the Ndc80 complex through the cell cycle. Cnn1 localizes to kinetochores at basal levels from G1 through metaphase but accumulates abruptly at anaphase onset. How Cnn1 is recruited and which activities regulate its dynamic localization are unclear. We show that Cnn1 harbors two kinetochore-localization activities: a C-terminal histone-fold domain (HFD) that associates with the centromere region and a N-terminal Spc24/Spc25 interaction sequence that mediates linkage to the microtubule-binding Ndc80 complex. We demonstrate that the established Ndc80 binding site in the N terminus of Cnn1, Cnn1 , should be extended with flanking residues, Cnn1 , to allow near maximal binding affinity to Ndc80. Cnn1 localization was proposed to depend on Mps1 kinase activity at Cnn1-S74, based on in vitro experiments demonstrating the Cnn1-Ndc80 complex interaction. We demonstrate that from G1 through metaphase, Cnn1 localizes via both its HFD and N-terminal Spc24/Spc25 interaction sequence, and deletion or mutation of either region results in anomalous Cnn1 kinetochore levels. At anaphase onset (when Mps1 activity decreases) Cnn1 becomes enriched mainly via the N-terminal Spc24/Spc25 interaction sequence. In sum, we provide the first in vivo evidence of Cnn1 preanaphase linkages with the kinetochore and enrichment of the linkages during anaphase. KEYWORDS Cnn1; Mps1; kinetochore; centromere; CENP-T K INETOCHORES are large protein structures that assemble hierarchically on the centromeres (CEN) of replicated chromosomes (sister chromatids). They biorient each sister chromatid pair to the microtubules (MTs) of the mitotic spindle and orchestrate chromatid segregation into the daughter cells (Cheeseman 2014;Malvezzi and Westermann 2014). At the core of each kinetochore lies a protein network named KMN (KLN1/Spc105, MIS12/Mtw1, and NDC80/Ndc80 complexes) that bridges the CEN and MTs (Cheeseman et al. 2006;Westermann et al. 2007). The Ndc80 complex attaches kinetochores to the MTs via its outer Ndc80/Nuf2 dimer (Wei et al. 2007;Ciferri et al. 2008;Alushin et al. 2010), while its CEN-proximal Spc24/Spc25 dimer interacts with a putatively CEN-associated protein, known as Cnn1 in budding yeast (CENP-T in metazoans). The N-terminal domain of Cnn1 and CENP-T hook onto the interface of the Spc24/Spc25 dimer (Malvezzi et al. 2013;Nishino et al. 2013). In its C terminus, Cnn1 harbors a histonefold domain (HFD) (Schleiffer et al. 2012), which may associate with CEN DNA, as does CENP-T (Hori et al. 2008;Nishino et al. 2012).Cnn1 levels at kinetochores are low from G1 through metaphase but increase two-to threefold at anaphase entry and drop back to base level at anaphase exit. Cnn1 interacts with the...

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Section: Discussionmentioning

confidence: 99%

The Mps1 Kinase Modulates the Recruitment and Activity of Cnn1CENP-T at Saccharomyces cerevisiae Kinetochores

et al. 2015

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“…CENP-A nucleosomes are interspersed with H3.1-containing nucleosomes at the inner region of centromeres (Blower et al, 2002;Ribeiro et al, 2010;Sullivan and Karpen, 2004). CENP-A is required for the assembly of a large multi-protein complex, the kinetochore, which forms the interface between chromatin and the spindle (Westhorpe and Straight, 2013). Loss of CENP-A results in errors during chromosome congression and segregation, which are a source of genomic instability (Régnier et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

“…In order to achieve chromosomal segregation, microtubules emanating from the spindle are bound to chromosomes at specific genomic loci called centromeres (Westhorpe and Straight, 2013). Spindle dynamics then drive chromosome movement and separation (Walczak et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

CRL4RBBP7 is required for efficient CENP-A deposition at centromeres

Mouysset¹,

Gilberto²,

Meier³

et al. 2015

Journal of Cell Science

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The mitotic spindle drives chromosome movement during mitosis and attaches to chromosomes at dedicated genomic loci named centromeres. Centromeres are epigenetically specified by their histone composition, namely the presence of the histone H3 variant CENP-A, which is regulated during the cell cycle by its dynamic expression and localization. Here, we combined biochemical methods and quantitative imaging approaches to investigate a new function of CUL4-RING E3 ubiquitin ligases (CRL4) in regulating CENP-A dynamics. We found that the core components CUL4 and DDB1 are required for centromeric loading of CENP-A, but do not influence CENP-A maintenance or pre-nucleosomal CENP-A levels. Interestingly, we identified RBBP7 as a substrate-specific CRL4 adaptor required for this process, in addition to its role in binding and stabilizing soluble CENP-A. Our data thus suggest that the CRL4 complex containing RBBP7 (CRL4 RBBP7 ) might regulate mitosis by promoting ubiquitin-dependent loading of newly synthesized CENP-A during the G1 phase of the cell cycle.

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“…Kinetochores that consist of more than 15 centromere proteins (CENPs) are vital for accurate chromosome segregation by mediating microtubule attachment and controlling the movement of chromosomes during mitosis and meiosis (6). Inappropriate expression of CENPs may be a major cause of CIN and tumorigenesis (7).…”

Section: Introductionmentioning

confidence: 99%

CENP-H regulates the cell growth of human hepatocellular carcinoma cells through the mitochondrial apoptotic pathway

Hou

et al. 2017

Oncology Reports

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Abstract. The genomic alterations of hepatocellular carcinoma (HCC) are still unclear. Centromere protein-H (CENP-H) has been shown to be associated with many solid tumors. Our previous study found that CENP-H was upregulated in HCC and was related to patient prognosis. However, the biological functions of CENP-H in HCC and the possible underlying mechanisms have not been well elucidated. In the present study, we demonstrated that CENP-H knockdown inhibited the proliferation of Hep3B cells and decreased colony formation ability of single cells in vitro. Furthermore, CENP-H knockdown induced Hep3B cell apoptosis, and apoptotic bodies were observed using transmission electron microscopy. The protein expression of cleaved caspase-3 was upregulated in Hep3B cells after CENP-H knockdown. Additionally, a Bax/Bcl-2 ratio imbalance with a significant increase of Bax and a substantial decrease of Bcl-2 at both the mRNA and protein levels were determined in this study. In an animal experiment, CENP-H knockdown blocked the growth of Hep3B subcutaneous xenografts. Immunohistochemistry revealed that the protein expression of cleaved caspase-3 and Bax was increased, whereas the protein expression of Bcl-2 and Ki-67 was decreased in subcutaneous xenografts of the CENP-H-knockdown group. In summary, CENP-H may be involved in cell proliferation and apoptosis of HCC cells through the mitochondrial apoptotic pathway. Combined with previous studies, the data provide a new perspective on HCC development and progression.

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Functions of the centromere and kinetochore in chromosome segregation

Cited by 113 publications

References 61 publications

The Mps1 Kinase Modulates the Recruitment and Activity of Cnn1CENP-T at Saccharomyces cerevisiae Kinetochores

The Mps1 Kinase Modulates the Recruitment and Activity of Cnn1CENP-T at Saccharomyces cerevisiae Kinetochores

CRL4RBBP7 is required for efficient CENP-A deposition at centromeres

CENP-H regulates the cell growth of human hepatocellular carcinoma cells through the mitochondrial apoptotic pathway

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