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Published 6 January 2003. doi:10.1083/jcb.200210005
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© The Rockefeller University Press, 0021-9525/2003/1/25 $5.00
The Journal of Cell Biology, Volume 160, Number 1, 25-39

Article

Human centromere chromatin protein hMis12, essential for equal segregation, is independent of CENP-A loading pathway



Gohta Goshima1, Tomomi Kiyomitsu1, Kinya Yoda2 and Mitsuhiro Yanagida1

1 COE Research Project, Department of Gene Mechanisms, Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan
2 Bioscience Center, Nagoya University, Nagoya 464-8601, Japan

Address correspondence to Mitsuhiro Yanagida, Department of Gene Mechanisms, Graduate School of Biostudies, Kyoto University, Kitashirakawa-Oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan. Tel.: 81-75-753-4205. Fax: 81-75-753-4208. E-mail: yanagida{at}kozo.biophys.kyoto-u.ac.jp

Kinetochores are the chromosomal sites for spindle interaction and play a vital role for chromosome segregation. The composition of kinetochore proteins and their cellular roles are, however, poorly understood in higher eukaryotes. We identified a novel kinetochore protein family conserved from yeast to human that is essential for equal chromosome segregation. The human homologue hMis12 of yeast spMis12/scMtw1 retains conserved sequence features and locates at the kinetochore region indistinguishable from CENP-A, a centromeric histone variant. RNA interference (RNAi) analysis of HeLa cells shows that the reduced hMis12 results in misaligned metaphase chromosomes, lagging anaphase chromosomes, and interphase micronuclei without mitotic delay, while CENP-A is located at kinetochores. Further, the metaphase spindle length is abnormally extended. Spindle checkpoint protein hMad2 temporally localizes at kinetochores at early mitotic stages after RNAi. The RNAi deficiency of CENP-A leads to a similar mitotic phenotype, but the kinetochore signals of other kinetochore proteins, hMis6 and CENP-C, are greatly diminished. RNAi for hMis6, like that of a kinetochore kinesin CENP-E, induces mitotic arrest. Kinetochore localization of hMis12 is unaffected by CENP-A RNAi, demonstrating an independent pathway of CENP-A in human kinetochores.

Key Words: kinetochore; HeLa; RNAi; Mis12; CENP-A


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