The distinct functions of CENP-C and CENP-T/W in centromere propagation and function in<i>Xenopus</i>egg extracts

Krizaic, Iva; Williams, Samantha; Sánchez, Patricia; Rodríguez-Corsino, Miriam; Stukenberg, P. Todd; Losada, Ana

doi:10.1080/19491034.2014.1003509

Cited by 16 publications

(29 citation statements)

References 53 publications

(89 reference statements)

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“…Strikingly, in extracts partially depleted of CENP-C, CENP-C levels were ~50% at kinetochores (Figures 6A, 6B and 6C), and BubR1 levels remained high in nocodazole but decreased dramatically upon treatment with STLC, compared to controls (Figures 6A, 6B and 6D). Full depletion of CENP-C, which caused a significant loss of outer kinetochore Ndc80 (Figures 6A, 6B and 6E) (Krizaic et al, 2015; Milks et al, 2009), resulted in a decrease in BubR1 in both conditions (Figure 6D). We conclude that complete assembly of CENP-C at kinetochores is specifically required for the enrichment of BubR1 at tensionless attachments.…”

Section: Resultsmentioning

confidence: 99%

Distinct Roles of the Chromosomal Passenger Complex in the Detection of and Response to Errors in Kinetochore-Microtubule Attachment

et al. 2017

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Summary The Chromosomal Passenger Complex (CPC) localizes to centromeres in early mitosis to activate its subunit Aurora B kinase. However, it is unclear if centromeric CPC localization contributes to CPC functions beyond Aurora B activation. Here, we show that an activated CPC that cannot localize to centromeres supports functional assembly of the outer kinetochore, but is unable to correct errors in kinetochore-microtubule attachment in Xenopus egg extracts. We find that CPC has two distinct roles at centromeres: one to properly phosphorylate Ndc80 to regulate attachment, and a second, conserved kinase-independent role in the proper composition of inner kinetochore proteins. Although a fully assembled inner kinetochore is not required for outer kinetochore assembly, we find it is essential to recruit tension indicators, such as BubR1 and 3F3/2, to erroneous attachments. Thus, centromeric CPC is necessary for tension-dependent removal of erroneous attachments, and for the kinetochore composition required to detect tension loss.

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Section: Resultsmentioning

confidence: 99%

Distinct Roles of the Chromosomal Passenger Complex in the Detection of and Response to Errors in Kinetochore-Microtubule Attachment

et al. 2017

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“…CENP-C appears to be upstream of all other CCAN complexes, including the CENP-H group in human cells (Klare et al 2015;Tachiwana et al 2015). Studies using Xenopus egg extract showed that CENP-C is the first CCAN component recruited to the centromere of sperm chromatin alongside new CENP-A (Milks et al 2009;Krizaic et al 2015). This suggests that CENP-C functions upstream of other CCAN components during de novo CCAN recruitment.…”

Section: Cenp-c Functions Upstream Of Other Ccan Components During Dementioning

confidence: 88%

“…2a and 3a). This is seen most clearly in germ line cells in Xenopus (Milks et al 2009;Krizaic et al 2015) and Drosophila (Raychaudhuri et al 2012), where CENP-A provides a template for fertilized zygote centromeres. In addition, disruption of the original centromere leads to efficient neocentromere formation in the regions surrounding the original centromere.…”

Section: Priming the Centromerementioning

confidence: 92%

“…However, CENP-T localization is not completely lost in CENP-H-deficient DT40 cells (Hori et al 2008a). CENP-T expression in Xenopus was not completely lost after depletion of CENP-C (Krizaic et al 2015). CENP-T-W-S-X contains histone fold domains, which confer an innate DNA-binding ability (Hori et al 2008a;Nishino et al 2012;Takeuchi et al 2014).…”

Section: Localization Of Cenp-t-w-s-x Depends On Its Dna-binding Actimentioning

confidence: 95%

“…These observations suggest that direct DNA interaction and the CENP-H group are both required for centromere localization of CENP-T-W-S-X. Experiments in Xenopus showed that de novo CENP-T recruitment occurs during S phase, but it is not coupled with DNA replication (Krizaic et al 2015). FRAP experiments using human cells suggest that CENP-T recruitment occurs in S phase (Prendergast et al 2011;Dornblut et al 2014).…”

Section: Localization Of Cenp-t-w-s-x Depends On Its Dna-binding Actimentioning

confidence: 96%

See 2 more Smart Citations

Kinetochore assembly and function through the cell cycle

Nagpal

Fukagawa

2016

Chromosoma

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The kinetochore is an essential structure for the chromosome segregation machinery in eukaryotes; it serves as a bridge between the spindle microtubules and chromosomes. The kinetochore consists of multiple interconnecting components on the centromere; therefore, understanding its formation, molecular function, and regulation has remained an ongoing challenge. Recent studies have provided new insights into centromere identity, kinetochore assembly, and function. In this review, we discuss recent advances in our understanding of the function and regulation of key kinetochore components. We highlight the reciprocal localization dependencies of the different sub-complexes of the kinetochore and describe their regulation during the cell cycle.

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The Power of Xenopus Egg Extract for Reconstitution of Centromere and Kinetochore Function

French

Straight

2017

Centromeres and Kinetochores

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Faithful transmission of genetic information during cell division requires attachment of chromosomes to the mitotic spindle via the kinetochore. In vitro reconstitution studies are beginning to uncover how the kinetochore is assembled upon the underlying centromere, how the kinetochore couples chromosome movement to microtubule dynamics, and how cells ensure the site of kinetochore assembly is maintained from one generation to the next. Here we give special emphasis to advances made in Xenopus egg extract, which provides a unique, biochemically tractable in vitro system that affords the complexity of cytoplasm and nucleoplasm to permit reconstitution of the dynamic, cell cycle-regulated functions of the centromere and kinetochore.

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

Cited by 16 publications

References 53 publications

Distinct Roles of the Chromosomal Passenger Complex in the Detection of and Response to Errors in Kinetochore-Microtubule Attachment

Distinct Roles of the Chromosomal Passenger Complex in the Detection of and Response to Errors in Kinetochore-Microtubule Attachment

Kinetochore assembly and function through the cell cycle

The Power of Xenopus Egg Extract for Reconstitution of Centromere and Kinetochore Function

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