Molecular mechanisms of kinetochore capture by spindle microtubules

Tanaka, Kozo; Mukae, Naomi; Dewar, Hilary; Breugel, M. van; James, Euan K.; Prescott, Alan R.; Antony, Claude; Tanaka, Tomoyuki

doi:10.1038/nature03483

Cited by 264 publications

(419 citation statements)

References 49 publications

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“…The original search-and-capture model supposed that KC attachments to MTs are end-on, consistent with the results of Kalinina et al (24), while others observed primarily lateral attachments (14,19,20,23,25). Therefore, we studied how the probability of lateral or end-on attachment varied with MT length and the presence or absence of rotational diffusion in the model (Fig.…”

Section: Discussionsupporting

confidence: 80%

Contributions of Microtubule Dynamic Instability and Rotational Diffusion to Kinetochore Capture

Blackwell

Sweezy-Schindler

Edelmaier

et al. 2017

Biophysical Journal

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Microtubule dynamic instability allows search and capture of kinetochores during spindle formation, an important process for accurate chromosome segregation during cell division. Recent work has found that microtubule rotational diffusion about minus-end attachment points contributes to kinetochore capture in fission yeast, but the relative contributions of dynamic instability and rotational diffusion are not well understood. We have developed a biophysical model of kinetochore capture in small fission-yeast nuclei using hybrid Brownian dynamics/kinetic Monte Carlo simulation techniques. With this model, we have studied the importance of dynamic instability and microtubule rotational diffusion for kinetochore capture, both to the lateral surface of a microtubule and at or near its end. Over a range of biologically relevant parameters, microtubule rotational diffusion decreased capture time, but made a relatively small contribution compared to dynamic instability. At most, rotational diffusion reduced capture time by 25%. Our results suggest that while microtubule rotational diffusion can speed up kinetochore capture, it is unlikely to be the dominant physical mechanism for typical conditions in fission yeast. In addition, we found that when microtubules undergo dynamic instability, lateral captures predominate even in the absence of rotational diffusion. Counterintuitively, adding rotational diffusion to a dynamic microtubule increases the probability of endon capture.

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Section: Discussionsupporting

confidence: 80%

Contributions of Microtubule Dynamic Instability and Rotational Diffusion to Kinetochore Capture

Blackwell

Sweezy-Schindler

Edelmaier

et al. 2017

Biophysical Journal

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“…In prometaphase, kinetochores, a protein structure formed at the centromere of each sister chromatid, 'capture' and interact with microtubules projecting from centrosomes (search and capture model) (Rieder and Salmon, 1998;Hayden et al, 1990;Tanaka et al, 2005;Tanaka et al, 2008). Kinetochores are then moved along the length of the attached microtubules toward the spindle pole promoted by kinetochore associated minus-end motor proteins (dynein in metazoans and kinesin-14 in budding yeast) (Rieder and Alexander, 1990;Yang et al, 2007b;King et al, 2000;Tanaka et al, 2007).…”

Section: Centrosome Sister Chromatidmentioning

confidence: 99%

An ATM- and ATR-dependent checkpoint inactivates spindle assembly by targeting CEP63

Smith¹,

Dejsuphong

Balestrini³

et al. 2009

Nat Cell Biol

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The effects of ATM and ATR signalling induced by chromosomal breakage have been described extensively in modulating cell cycle progression up to the onset of mitosis.However, DNA damage checkpoint responses in mitotic cells are not well understood.This thesis reports on the effects of double strand breaks on the progression of mitosis.We found ATM and ATR activation can occur in mitotic Xenopus laevis egg extract and the induction of ATM and ATR by chromosomal breakages inhibits spindle assembly in both Xenopus egg extract and somatic cells. The delay in mitotic progression induced by ATM and ATR was found not to involve major spindle assembly factors activities such as, Cdk1, Plx1 and RCC1/Ran-GTP. However, normal anastral spindles formation around linear DNA coated beads, which can activate ATM and ATR, linked centrosome-driven spindle assembly to ATM and ATR dependent spindle defects. cDNA expression library screening was undertaken to identify novel ATM and ATR targets in this mitotic checkpoint pathway, through which the novel centrosomal protein XCEP63 was identified as a likely candidate. Data obtained from depletion and reconstitution of XCEP63 in Xenopus egg extract established that normal centrosome-driven spindle assembly requires XCEP63. Moreover, ATM and ATR phosphorylates XCEP63 on serine 560 and promotes delocalisation from the centrosome. ATM and ATR inhibition or addition of non-phosphorylable XCEP63 recombinant protein mutated at serine 560 prevents spindle assembly abnormalities.These findings suggest that ATM and ATR regulate mitotic events by targeting XCEP63 and centrosome-dependent spindle assembly. This pathway may provide support for DNA repair processes or regulate cell survival in the presence of mitotic DNA damage. 3

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“…We were not able to analyze the main-tenance of a previously assembled kinetochore because cells needed to pass through S phase to deplete Cse4 from the eCEN. Because kinetochores likely disassemble during S phase (Tanaka et al, 2005), studies at the eCEN would not distinguish between kinetochore assembly and maintenance. All proteins tested exhibited some dependence on Cse4 for localization to the cCEN.…”

Section: The Functions Of Cenh3mentioning

confidence: 99%

“…When cells are grown in galactose media, transcription through the cCEN keeps it inactive. The addition of glucose represses transcription at the cCEN, allowing a functional kinetochore to form within 20 min (Hill and Bloom, 1989;Neff and Burke, 1992;Dewar et al, 2004;Tanaka et al, 2005).To begin analyzing the cell cycle restrictions over kinetochore assembly, we first determined whether kinetochores could assemble in G 1 cells. Kinetochore assembly at the cCEN was analyzed using ChIP.…”

mentioning

confidence: 99%

De Novo Kinetochore Assembly Requires the Centromeric Histone H3 Variant

Collins

Castillo

Tatsutani

et al. 2005

MBoC

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Kinetochores mediate chromosome attachment to the mitotic spindle to ensure accurate chromosome segregation. Budding yeast is an excellent organism for kinetochore assembly studies because it has a simple defined centromere sequence responsible for the localization of >65 proteins. In addition, yeast is the only organism where a conditional centromere is available to allow studies of de novo kinetochore assembly. Using a conditional centromere, we found that yeast kinetochore assembly is not temporally restricted and can occur in both G 1 phase and prometaphase. We performed the first investigation of kinetochore assembly in the absence of the centromeric histone H3 variant Cse4 and found that all proteins tested depend on Cse4 to localize. Consistent with this observation, Cse4-depleted cells had severe chromosome segregation defects. We therefore propose that yeast kinetochore assembly requires both centromeric DNA specificity and centromeric chromatin. INTRODUCTIONAccurate chromosome segregation in mitosis and meiosis is essential for the maintenance of genomic stability. Chromosomes attach to the mitotic spindle at the kinetochore, the protein complex that assembles onto centromeric DNA. Although kinetochore function is conserved, the underlying centromeric DNA is highly variable. Budding yeast contain a 125-base pair sequence-specific centromere that is sufficient for kinetochore formation (Fitzgerald-Hayes et al., 1982). In contrast, centromeres in multicellular eukaryotes are composed of megabases of highly repetitive DNA that lack sequence specificity (for review, see Sullivan et al., 2001). In these organisms, kinetochore assembly seems to be propagated by unidentified epigenetic component(s) (Karpen and Allshire, 1997;Sullivan et al., 2001).The best-characterized kinetochore is in budding yeast where Ͼ65 components have been identified that constitutively localize to the kinetochore (for reviews, see Biggins and Walczak, 2003;McAinsh et al., 2003). Most of the yeast kinetochore proteins are found in biochemically distinct complexes known as the CBF3, CTF19/COMA, MTW1, NDC80, and DAM1 complexes that seem to assemble on a single centromeric nucleosome (Meluh et al., 1998). Although the exact architecture of the kinetochore is not known, dependency relationships subdivide the kinetochore into inner, central, and outer domains. The inner kinetochore contains the CBF3 complex (Ndc10, Cep3, Skp1, and Ctf13) as well as the DNA binding proteins Mif2, Cbf1, and the yeast centromeric histone H3 variant (CenH3) Cse4. CBF3 binds directly to the centromeric DNA and is thought to nucleate kinetochore assembly because all kinetochore proteins require it for localization (Russell et al., 1999;Goshima and Yanagida, 2000;He et al., 2001;Janke et al., 2001Janke et al., , 2002. The central kinetochore contains the MTW1 (Mtw1, Dsn1, Nnf1, and Nsl1) and CTF19/COMA (Ctf19, Mcm16, Mcm19, Mcm21, Mcm22, Ctf3, Chl4, Okp1, Ame1, Iml3, Nkp1, and Nkp2) complexes. CTF19/COMA can be further divided into two subcomplexes, with Ame1...

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Molecular mechanisms of kinetochore capture by spindle microtubules

Cited by 264 publications

References 49 publications

Contributions of Microtubule Dynamic Instability and Rotational Diffusion to Kinetochore Capture

Contributions of Microtubule Dynamic Instability and Rotational Diffusion to Kinetochore Capture

An ATM- and ATR-dependent checkpoint inactivates spindle assembly by targeting CEP63

De Novo Kinetochore Assembly Requires the Centromeric Histone H3 Variant

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