Molecular Analysis of Kinetochore-Microtubule Attachment in Budding Yeast

He, Xiangwei; Rines, Daniel R.; Espelin, Christopher W.; Sorger, Peter K.

doi:10.1016/s0092-8674(01)00438-x

Cited by 231 publications

(310 citation statements)

References 54 publications

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“…However, by performing immunofluorescence on chromosome spreads of insoluble nuclear material, we were able to detect the kinetochore localization of Ipl1p at metaphase, suggesting that Ipl1p is in the Aurora B family. Our localization results are similar to those recently published (He et al 2001). Finding Ipl1p at kinetochores is consistent with its role in the spindle checkpoint.…”

Section: Ipl1p Localizes To Kinetochores During Metaphasesupporting

confidence: 92%

The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint

Biggins¹,

Murray²

2001

Genes Dev.

391

395

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The spindle checkpoint prevents cell cycle progression in cells that have mitotic spindle defects. Although several spindle defects activate the spindle checkpoint, the exact nature of the primary signal is unknown. We have found that the budding yeast member of the Aurora protein kinase family, Ipl1p, is required to maintain a subset of spindle checkpoint arrests. Ipl1p is required to maintain the spindle checkpoint that is induced by overexpression of the protein kinase Mps1. Inactivating Ipl1p allows cells overexpressing Mps1p to escape from mitosis and segregate their chromosomes normally. Therefore, the requirement for Ipl1p in the spindle checkpoint is not a consequence of kinetochore and/or spindle defects. The requirement for Ipl1p distinguishes two different activators of the spindle checkpoint: Ipl1p function is required for the delay triggered by chromosomes whose kinetochores are not under tension, but is not required for arrest induced by spindle depolymerization. Ipl1p localizes at or near kinetochores during mitosis, and we propose that Ipl1p is required to monitor tension at the kinetochore. The accurate propagation of genetic information depends on faithful chromosome segregation. Accurate chromosome segregation depends on the precise coordination of events in the chromosome cycle. When chromosomes replicate during S phase, linkage between the sister chromatids (cohesion) is established and must be maintained while chromosomes condense and align on the mitotic spindle. Chromosomes attach to the mitotic spindle by their kinetochores, specialized protein structures that are assembled on centromeric DNA sequences. Once all the chromosomes are correctly aligned on the mitotic spindle, sister chromatid cohesion must dissolve promptly at anaphase to allow the sister chromatids to segregate rapidly to opposite poles of the mitotic spindle. Defects in any of these steps can result in aneuploidy, a hallmark of tumor cells and some birth defects (Lengauer et al. 1997.The spindle checkpoint prevents cells from separating their sister chromatids until chromosome alignment is complete. The conserved components of the checkpoint include the Mad (Mad1-Mad3) proteins, Bub1 and Bub3, Mps1 (a protein kinase), and Cdc55 (Hoyt et al. 1991;Li and Murray 1991;Minshull et al. 1996;Weiss and Winey 1996;Wang and Burke 1997). A separate control, the Bub2-dependent checkpoint, monitors a second aspect of chromosome segregation, the delivery of DNA or a spindle pole body into the daughter cell (Alexandru et al. 1999;Fesquet et al. 1999;Fraschini et al. 1999;Li 1999). Spindle checkpoint defects are associated with genetic instability, and some human cancers contain mutant spindle checkpoint genes (Cahill et al. 1998;Takahashi et al. 1999) The spindle checkpoint monitors the interaction between kinetochores and microtubules. Spindle checkpoint proteins localize to kinetochores (Chen et al. 1996;Taylor and McKeon 1997;Bernard et al. 1998), and all known kinetochore and spindle defects that activate the checkpoint affect the i...

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Section: Ipl1p Localizes To Kinetochores During Metaphasesupporting

confidence: 92%

The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint

Biggins¹,

Murray²

2001

Genes Dev.

391

395

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“…However, in contrast to studies that suggested the existence of CenH3-independent assembly pathways based on immunofluorescence data (Oegema et al, 2001;Goshima et al, 2003;Hayashi et al, 2004;Regnier et al, 2005), we found that all proteins tested exhibited some dependence on Cse4 when the ChIP technique was used to quantify occupancy. Although the inner kinetochore complex CBF3 was previously proposed to nucleate yeast kinetochore assembly (Russell et al, 1999;Goshima and Yanagida, 2000;He et al, 2001;Janke et al, 2001Janke et al, , 2002, our work shows that the centromeric nucleosome also has an essential role in kinetochore assembly. We therefore favor a model where the centromeric nucleosome creates a chromatin structure that cooperates with the binding of the inner CBF3 complex to initiate yeast kinetochore assembly.…”

Section: The Functions Of Cenh3mentioning

confidence: 54%

“…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.…”

Section: Introductionmentioning

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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“…Recent data has suggested that knock down of one of these three proteins can cause concomittant reduction in the expression of other complex members. 10,14 The emerging view is that within the chromosomal passenger complex, aurora B kinase is the active component responsible for regulating biorientation of chromosomes during prometaphase, [15][16][17] while INCENP, survivin and borealin are thought to direct aurora B to its substrates and possibly enhance its kinase activity. 9,10,14,[18][19][20][21] Aurora B kinase has an ever growing list of substrates, which includes itself, 22 its fellow passengers, borealin, 12 INCENP, 18 and survivin 23 and a variety of other chromosomal associated proteins such as histone 3, 24 the kinI kinesin, MCAK [25][26][27] and topoisomerase II; 28 for reviews see refs.…”

Section: Introductionmentioning

confidence: 99%