Comprehensive analysis of the ICEN (Interphase Centromere Complex) components enriched in the CENP‐A chromatin of human cells

Izuta, Hiroshi; Ikeno, Mitsuru; Suzuki, Nobutaka; Tomonaga, Takeshi; Nozaki, Naohito; Obuse, Chikashi; Kisu, Yasutomo; Goshima, Naoki; Nomura, Fumio; Nomura, Nobuo; Yoda, Kinya

doi:10.1111/j.1365-2443.2006.00969.x

Cited by 193 publications

(185 citation statements)

References 36 publications

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“…CENP-K depletion reduced the kinetochore levels of CENP-C in agreement with prior analysis of the effect of CENP-H and -K depletions on CENP-C localization in interphase cells (Fukagawa et al, 2001;Kwon et al, 2007) and with the close association of CENP-H/I/K with CENP-C (Foltz et al, 2006;Izuta et al, 2006; Figure 2A). Because CENP-H and CENP-Q localization is strongly affected after CENP-K RNAi ( Figure 2A; also see quantitative analysis in Okada et al, 2006), this suggests that the RNAi conditions used here do significantly reduce CENP-K levels.…”

Section: Knl1 Does Not Contribute To Localization Of Constitutive Censupporting

confidence: 76%

KNL1 and the CENP-H/I/K Complex Coordinately Direct Kinetochore Assembly in Vertebrates

Cheeseman

Hori

Fukagawa

et al. 2008

MBoC

172

188

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Chromosome segregation during mitosis requires the assembly of a large proteinaceous structure termed the kinetochore. In Caenorhabditis elegans, KNL-1 is required to target multiple outer kinetochore proteins. Here, we demonstrate that the vertebrate KNL1 counterpart is essential for chromosome segregation and is required to localize a subset of outer kinetochore proteins. However, unlike in C. elegans, depletion of vertebrate KNL1 does not abolish kinetochore localization of the microtubule-binding Ndc80 complex. Instead, we show that KNL1 and CENP-K, a subunit of a constitutively centromere-associated complex that is missing from C. elegans, coordinately direct Ndc80 complex localization. Simultaneously reducing both hKNL1 and CENP-K function abolishes all aspects of kinetochore assembly downstream of centromeric chromatin and causes catastrophic chromosome segregation defects. These findings explain discrepancies in kinetochore assembly pathways between different organisms and reveal a surprising plasticity in the assembly mechanism of an essential cell division organelle.

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Section: Knl1 Does Not Contribute To Localization Of Constitutive Censupporting

confidence: 76%

KNL1 and the CENP-H/I/K Complex Coordinately Direct Kinetochore Assembly in Vertebrates

Cheeseman

Hori

Fukagawa

et al. 2008

MBoC

172

188

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“…In recent years, many kinetochore proteins have been identified in vertebrate cells by various approaches (Cheeseman et al, 2004;Obuse et al, 2004a,b;Minoshima et al, 2005;Foltz et al, 2006;Izuta et al, 2006;McAinsh et al, 2006;Meraldi et al, 2006;Okada et al, 2006). Characterization of the cellular functions of each of these proteins and the protein-protein network within cells will lead to an understanding of how kinetochores assemble and function.…”

Section: Introductionmentioning

confidence: 99%

CENP-O Class Proteins Form a Stable Complex and Are Required for Proper Kinetochore Function

Hori

Okada

Maenaka

et al. 2008

MBoC

125

176

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We previously identified a multisubunit complex (CENP-H/I complex) in kinetochores from human and chicken cells. We showed that the CENP-H/I complex is divided into three functional classes. In the present study, we investigated CENP-O class proteins, which include CENP-O, -P, -Q, -R, and -50 (U). We created chicken DT40 cell knockouts of each of these proteins, and we found that all knockout lines were viable, but that they showed slow proliferation and mitotic defects. Kinetochore localization of CENP-O, -P, -Q, and -50 was interdependent, but kinetochore localization of these proteins was observed in CENP-R-deficient cells. A coexpression assay in bacteria showed that CENP-O, -P, -Q, and -50 proteins form a stable complex that can associate with CENP-R. Phenotype analysis of knockout cells showed that all proteins except for CENP-R were required for recovery from spindle damage, and phosphorylation of CENP-50 was essential for recovery from spindle damage. We also found that treatment with the proteasome inhibitor MG132 partially rescued the severe mitotic phenotype observed in response to release from nocodazole block in CENP-50 -deficient cells. This suggests that CENP-O class proteins are involved in the prevention of premature sister chromatid separation during recovery from spindle damage.

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“…The CCAN remains associated with centromeric chromatin across the entire cell cycle (6)(7)(8). The CCAN includes CENP-A, CENP-C, and four multisubunit complexes: CENP-L/-N (9), CENP-H/-I/-K/-M (10), CENP-O/-P/Q-/R/-U (11), and CENP-T/-W/-S/-X (12)(13)(14)(15).…”

mentioning

confidence: 99%

Stepwise unfolding supports a subunit model for vertebrate kinetochores

Vargiu

Макаров

Allan

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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During cell division, interactions between microtubules and chromosomes are mediated by the kinetochore, a proteinaceous structure located at the primary constriction of chromosomes. In addition to the centromere histone centromere protein A (CENP-A), 15 other members of the constitutive centromere associated network (CCAN) participate in the formation of a chromatin-associated scaffold that supports kinetochore structure. We performed a targeted screen analyzing unfolded centrochromatin from CENP-depleted chromosomes. Our results revealed that CENP-C and CENP-S are critical for the stable folding of mitotic kinetochore chromatin. Multipeak fitting algorithms revealed the presence of an organized pattern of centrochromatin packing consistent with arrangement of CENP-A-containing nucleosomes into up to five chromatin "subunits"-each containing roughly 20-30 nucleosomes. These subunits could be either layers of a boustrophedon or small loops of centromeric chromatin.centromere | kinetochore | CENP-A | centrochromatin | boustrophedon

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Comprehensive analysis of the ICEN (Interphase Centromere Complex) components enriched in the CENP‐A chromatin of human cells

Abstract: The centromere is a chromatin structure essential for correct segregation of sister chromatids, and defects in this region often lead to aneuploidy and cancer. We have previously reported purification of the interphase centromere complex (

Cited by 193 publications

References 36 publications

KNL1 and the CENP-H/I/K Complex Coordinately Direct Kinetochore Assembly in Vertebrates

KNL1 and the CENP-H/I/K Complex Coordinately Direct Kinetochore Assembly in Vertebrates

CENP-O Class Proteins Form a Stable Complex and Are Required for Proper Kinetochore Function

Stepwise unfolding supports a subunit model for vertebrate kinetochores

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