Stable Kinetochore-Microtubule Attachment Constrains Centromere Positioning in Metaphase

Pearson, Chad G.; Yeh, Elaine; Gardner, Melissa K.; Odde, David J.; Salmon, Edward D.; Bloom, Kerry

doi:10.1016/j.cub.2004.09.086

Cited by 148 publications

(182 citation statements)

References 27 publications

(1 reference statement)

Supporting

Mentioning

163

Contrasting

Order By: Relevance

“…Our previous work showed that STB is free of Cse4 during G1, or when the mitotic spindle is depolymerized (12). In contrast, CEN is occupied by Cse4 in G1-arrested cells or those lacking the spindle (12,27). We exploited these differences between CEN and STB in their association with Cse4 to further characterize the contribution of Cse4 to the topological status of STB.…”

Section: Positive Writhe Of Stb Chromatin Is Directly Correlated To Itsmentioning

confidence: 99%

Histone H3-variant Cse4-induced positive DNA supercoiling in the yeast plasmid has implications for a plasmid origin of a chromosome centromere

Huang

Chang

Cui

et al. 2011

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

View full text Add to dashboard Cite

The Saccharomyces cerevisiae 2-μm plasmid is a multicopy selfish genome that resides in the nucleus. The genetic organization of the plasmid is optimized for stable, high-copy propagation in hostcell populations. The plasmid's partitioning system poaches host factors, including the centromere-specific histone H3-variant Cse4 and the cohesin complex, enabling replicated plasmid copies to segregate equally in a chromosome-coupled fashion. We have characterized the in vivo chromatin topology of the plasmid partitioning locus STB in its Cse4-associated and Cse4-nonassociated states. We find that the occupancy of Cse4 at STB induces positive DNA supercoiling, with a linking difference (ΔLk) contribution estimated between +1 and +2 units. One plausible explanation for this contrary topology is the presence of a specialized Cse4-containing nucleosome with a right-handed DNA writhe at a functional STB, contrasted by a standard histone H3-containing nucleosome with a left-handed DNA writhe at a nonfunctional STB. The similarities between STB and centromere in their nucleosome signature and DNA topology would be consistent with the potential origin of the unusual point centromere of budding yeast chromosomes from the partitioning locus of an ancestral plasmid.CEN evolution | nucleosome topology | reversome T he 2-μm plasmid of Saccharomyces cerevisiae resides in the nucleus at 40 to 60 copies per cell, and propagates itself with nearly the same stability as chromosomes (1, 2). The plasmid is a benign selfish DNA element that seems to provide no advantage to its host, but poses, at its normal copy number, no serious disadvantage either. In haploid cells the plasmid is organized into a cluster of three to five foci, and segregates as a cluster (3). This effective reduction in copy number necessitates an active partitioning system, comprised of two plasmid-coded proteins, Rep1 and Rep2, and a cis-acting partitioning locus STB, to ensure equal or nearly equal plasmid segregation. A decline in plasmid copy number because of rare missegregation events is corrected via DNA amplification triggered by the Flp sitespecific recombination system harbored by the plasmid (4, 5). Positive and negative regulatory circuits implemented through the plasmid-coded Raf1 protein and the Rep proteins ensure quick amplification response without the danger of runaway increase in copy number (6-8).The Rep-STB system channels several host factors involved in chromosome segregation toward the execution of plasmid segregation. These factors include the mitotic spindle, the spindleassociated motor Kip1, the RSC2 chromatin-remodeling complex, the centromere-specific histone H3-variant Cse4 (CenH3), and the yeast cohesin complex (9-15). The de novo assembly of the plasmid-partitioning complex at STB during the G1-S window of each cell cycle (9,12,14,16) is reminiscent of the assembly of the kinetochore complex at centromeres. However, kinetochore components have not been detected at STB by ChIP (13).The assembly of the plasmid-partitioning complex culmin...

show abstract

Section: Positive Writhe Of Stb Chromatin Is Directly Correlated To Itsmentioning

confidence: 99%

Histone H3-variant Cse4-induced positive DNA supercoiling in the yeast plasmid has implications for a plasmid origin of a chromosome centromere

Huang

Chang

Cui

et al. 2011

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

View full text Add to dashboard Cite

show abstract

“…Cse4p, the budding yeast CENP-A homologue, and Nuf2p, an essential kinetochore protein involved in checkpoint monitoring of the kinetochore (35,36), form two foci that represent clusters of separated sister CENs in metaphase (37). These foci segregate to opposite SPBs during anaphase and remain proximal to the SPBs into the next cell cycle ( Fig.…”

Section: Kinetochore Proteins Are Highly Dynamic and Exhibit Programmedmentioning

confidence: 99%

The kinetochore protein Ndc10p is required for spindle stability and cytokinesis in yeast

Bouck

Bloom

2005

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

View full text Add to dashboard Cite

The budding yeast kinetochore is comprised of >60 proteins and associates with 120 bp of centromeric (CEN) DNA. Kinetochore proteins are highly dynamic and exhibit programmed cell cycle changes in localization. The CEN-specific histone, Cse4p, is one of a few stable kinetochore components and remains associated with CEN DNA throughout mitosis. In contrast, several other kinetochore proteins have been observed along interpolar microtubules and at the midzone during anaphase. The inner kinetochore protein, Ndc10p, is enriched at the spindle midzone in late anaphase. We show that Ndc10p is transported to the plus-ends of interpolar microtubules at the midzone during anaphase, a process that requires survivin (Bir1p), a member of the aurora kinase (Ipl1p) complex, and Cdc14p phosphatase. In addition, Ndc10p is required for essential non-kinetochore processes during mitosis. Cells lacking functional Ndc10p show defects in spindle stability during anaphase and failure to split the septin ring during cytokinesis. This latter phenotype leads to a cell separation defect in ndc10 -1 cells. We propose that Ndc10p plays a direct role in maintaining spindle stability during anaphase and coordinates the completion of cell division after chromosome segregation.microtubule ͉ survivin ͉ mitosis ͉ septin

show abstract

“…To do these experiments, we first investigated whether cells could be depleted of Cse4 at the eCEN. We hypothesized that cells would need to pass through S phase in the absence of Cse4 to achieve depletion at the eCEN because fluorescence recovery after photobleaching experiments demonstrated that Cse4 is replaced at the centromere during S phase (Pearson et al, 2004). To facilitate G 1 arrest and allow the analysis of cells at anaphase, Degron-CSE4 pGAL-UBR1-myc cells were also deleted for the MAD2 spindle checkpoint gene, which halts the cell cycle when there is a defect in proper spindle assembly (for review, see Lew and Burke, 2003).…”

Section: Cse4 Can Be Depleted From the Endogenous Centromerementioning

confidence: 99%

“…It is likely that assembly is coupled to centromere replication because yeast kinetochores quickly achieve bioriented attachments after centromere duplication and Cse4 deposition occurs at this time (Goshima and Yanagida, 2000;He et al, 2000;Pearson et al, 2004).…”

Section: Yeast Kinetochore Assembly Is Not Cell Cycle Restrictedmentioning

confidence: 99%

De Novo Kinetochore Assembly Requires the Centromeric Histone H3 Variant

Collins

Castillo

Tatsutani

et al. 2005

MBoC

View full text Add to dashboard Cite

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...

show abstract

Stable Kinetochore-Microtubule Attachment Constrains Centromere Positioning in Metaphase

Cited by 148 publications

References 27 publications

Histone H3-variant Cse4-induced positive DNA supercoiling in the yeast plasmid has implications for a plasmid origin of a chromosome centromere

Histone H3-variant Cse4-induced positive DNA supercoiling in the yeast plasmid has implications for a plasmid origin of a chromosome centromere

The kinetochore protein Ndc10p is required for spindle stability and cytokinesis in yeast

De Novo Kinetochore Assembly Requires the Centromeric Histone H3 Variant

Contact Info

Product

Resources

About