The centromeric nucleosome-like CENP–T–W–S–X complex induces positive supercoils into DNA

Takeuchi, K.; Nishino, Tatsuya; Mayanagi, Kouta; Horikoshi, Naoki; Osakabe, Akihisa; Tachiwana, Hiroaki; Hori, Tomohide; Kurumizaka, Hitoshi; Fukagawa, Tatsuo

doi:10.1093/nar/gkt1124

Cited by 72 publications

(82 citation statements)

References 27 publications

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“…Thus, CENP-T could be assembled first onto replicated chromatin and later on recruit CENP-W/S/X. In vitro assays suggest the existence of multiple DNA binding sites in CENP-T and CENP-W. 28,38 While in human cells the assembly of the CENP-T/W/S/X complex on chromatin occurs in late S/G2 phase cells, 18,36 it may take place in mitosis in the egg extracts. Xenopus homologues of CENP-S and CENP-X are present in the database but no antibodies against them are available yet to test when they are recruited to centromeres.…”

Section: Discussionmentioning

confidence: 99%

“…35,36 These two proteins together with CENP-S and CENP-X, all 4 containing histone-fold domains, have been proposed to form a nucleosome-like particle capable of supercoiling DNA in vitro. 37,38 How CENP-T/W/S/X are targeted to the centromere is currently unclear although it has been reported that their incorporation occurs in late S phase/G2 and requires CENP-A. 18,35,36,39 It is also unclear whether they contribute to CENP-A deposition.…”

Section: Introductionmentioning

confidence: 99%

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The distinct functions of CENP-C and CENP-T/W in centromere propagation and function inXenopusegg extracts

Krizaic

Williams

Sánchez

et al. 2015

Nucleus

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The distinct functions of CENP-C and CENP-T/W in centromere propagation and function inXenopusegg extracts

Krizaic

Williams

Sánchez

et al. 2015

Nucleus

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“…This is consistent with the more diffuse signal observed for Cnn1 DHFD -GFP ( Figure S5) compared to Cnn1-GFP. Although the CEN-HFD interaction has not been delineated in yeast, Cnn1 may form a nucleosome-like structure as suggested for CENP-T (Nishino et al 2012;Nishino et al 2013;Takeuchi et al 2014). However, Basilico et al (2014) proposed a nonnucleosomal population, first because the CENP-HIKM complex is required for CENP-T recruitment and second because CENP-T turns over at CENs (Prendergast et al 2011).…”

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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“…In vitro, a heterotetramer of CENP-T, -W, -S, and -X was proposed to protect 100 bp of DNA from nuclease digestion, in a similar manner to H3 and CENP-A nucleosomes (Nishino et al 2012). More recently, a CENP-T/W/S/X octamer has been shown to bind 100 bp of DNA and potentially induce positive supercoils (as opposed to negative supercoils induced by nucleosomes) (Takeuchi et al 2014). However, whether a CENP-T/W/S/X nucleosome-like species exists at centromeres remains unclear, as the path of DNAwrapping around these proteins is unknown and the nuclease protection pattern is distinct from that of a nucleosome.…”

Section: Histone Modificationsmentioning

confidence: 99%

The Centromere: Epigenetic Control of Chromosome Segregation during Mitosis

Westhorpe

Straight

2014

Cold Spring Harb Perspect Biol

140

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A fundamental challenge for the survival of all organisms is maintaining the integrity of the genome in all cells. Cells must therefore segregate their replicated genome equally during each cell division. Eukaryotic organisms package their genome into a number of physically distinct chromosomes, which replicate during S phase and condense during prophase of mitosis to form paired sister chromatids. During mitosis, cells form a physical connection between each sister chromatid and microtubules of the mitotic spindle, which segregate one copy of each chromatid to each new daughter cell. The centromere is the DNA locus on each chromosome that creates the site of this connection. In this review, we present a brief history of centromere research and discuss our current knowledge of centromere establishment, maintenance, composition, structure, and function in mitosis.

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The centromeric nucleosome-like CENP–T–W–S–X complex induces positive supercoils into DNA

Cited by 72 publications

References 27 publications

The distinct functions of CENP-C and CENP-T/W in centromere propagation and function inXenopusegg extracts

The distinct functions of CENP-C and CENP-T/W in centromere propagation and function inXenopusegg extracts

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

The Centromere: Epigenetic Control of Chromosome Segregation during Mitosis

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