Mobility of kinetochore proteins measured by FRAP analysis in living cells-Reference-Cited by-同舟云学术

Mobility of kinetochore proteins measured by FRAP analysis in living cells

Published:2022-01-08 Issue:1 Volume:30 Page:43-57
ISSN:0967-3849
Container-title:Chromosome Research
language:en
Short-container-title:Chromosome Res

Author:

Watanabe Reito^ORCID,Hirano Yasuhiro^ORCID,Hara Masatoshi^ORCID,Hiraoka Yasushi^ORCID,Fukagawa Tatsuo^ORCID

Abstract

AbstractThe kinetochore is essential for faithful chromosome segregation during mitosis and is assembled through dynamic processes involving numerous kinetochore proteins. Various experimental strategies have been used to understand kinetochore assembly processes. Fluorescence recovery after photobleaching (FRAP) analysis is also a useful strategy for revealing the dynamics of kinetochore assembly. In this study, we introduced fluorescence protein-tagged kinetochore protein cDNAs into each endogenous locus and performed FRAP analyses in chicken DT40 cells. Centromeric protein (CENP)-C was highly mobile in interphase, but immobile during mitosis. CENP-C mutants lacking the CENP-A-binding domain became mobile during mitosis. In contrast to CENP-C, CENP-T and CENP-H were immobile during both interphase and mitosis. The mobility of Dsn1, which is a component of the Mis12 complex and directly binds to CENP-C, depended on CENP-C mobility during mitosis. Thus, our FRAP assays provide dynamic aspects of how the kinetochore is assembled.

Publisher

Springer Science and Business Media LLC

Subject

Genetics

Link

https://link.springer.com/content/pdf/10.1007/s10577-021-09678-x.pdf

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