RCC1 regulates inner centromeric composition in a Ran-independent fashion

Cell Cycle
Michael Shaofei ZhangMary Dasso

Abstract

RCC1 associates to chromatin dynamically within mitosis and catalyzes Ran-GTP production. Exogenous RCC1 disrupts kinetochore structure in Xenopus egg extracts (XEEs), but the molecular basis of this disruption remains unknown. We have investigated this question, utilizing replicated chromosomes that possess paired sister kinetochores. We find that exogenous RCC1 evicts a specific subset of inner KT proteins including Shugoshin-1 (Sgo1) and the chromosome passenger complex (CPC). We generated RCC1 mutants that separate its enzymatic activity and chromatin binding. Strikingly, Sgo1 and CPC eviction depended only on RCC1's chromatin affinity but not its capacity to produce Ran-GTP. RCC1 similarly released Sgo1 and CPC from synthetic kinetochores assembled on CENP-A nucleosome arrays. Together, our findings indicate RCC1 regulates kinetochores at the metaphase-anaphase transition through Ran-GTP-independent displacement of Sgo1 and CPC.

References

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Dec 3, 2014·Developmental Cell·Michael Shaofei ZhangMary Dasso
Nov 14, 2015·Molecular Biology of the Cell·Maiko FurutaTatsuo Fukagawa

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Citations

Aug 8, 2020·Biology of the Cell·Gabriel Guilloux, Romain Gibeaux
Jun 3, 2018·International Journal of Molecular Sciences·Huiying LiNan Zheng

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Methods Mentioned

BETA
nucleotide exchange
immunodepletion
gel filtration
Infrared Imaging

Software Mentioned

AlphaView
Prism

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