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Published online
doi:10.1083/jcb.200903088
The Journal of Cell Biology, Vol. 185, No. 3, 397-407
The Rockefeller University Press, 0021-9525 $30.00
© Perpelescu et al.
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Article

Active establishment of centromeric CENP-A chromatin by RSF complex



Marinela Perpelescu1, Naohito Nozaki2, Chikashi Obuse3, Hua Yang1, and Kinya Yoda1

1 Bioscience and Biotechnology Center, Nagoya University, Nagoya, Aichi 464-8601, Japan
2 Kanagawa Dental College, Yokosuka, Kanagawa 238-8580, Japan
3 Faculty of Advanced Life Science, Hokkaido University, Sapporo, Hokkaido 001-0021, Japan

Correspondence to Kinya Yoda: i45156a{at}cc.nagoya-u.ac.jp; or yodakinya{at}mac.com

Centromeres are chromosomal structures required for equal DNA segregation to daughter cells, comprising specialized nucleosomes containing centromere protein A (CENP-A) histone, which provide the basis for centromeric chromatin assembly. Discovery of centromere protein components is progressing, but knowledge related to their establishment and maintenance remains limited. Previously, using anti-CENP-A native chromatin immunoprecipitation, we isolated the interphase–centromere complex (ICEN). Among ICEN components, subunits of the remodeling and spacing factor (RSF) complex, Rsf-1 and SNF2h proteins, were found. This paper describes the relationship of the RSF complex to centromere structure and function, demonstrating its requirement for maintenance of CENP-A at the centromeric core chromatin in HeLa cells. The RSF complex interacted with CENP-A chromatin in mid-G1. Rsf-1 depletion induced loss of centromeric CENP-A, and purified RSF complex reconstituted and spaced CENP-A nucleosomes in vitro. From these data, we propose the RSF complex as a new factor actively supporting the assembly of CENP-A chromatin.

M. Perpelescu's present address is Department of Molecular Genetics, Mishima, Shizuoka, 411-8540, Japan.

Abbreviations used in this paper: CENP, centromere protein; ICEN, interphase–centromere complex; nChIP, native chromatin immunoprecipitation; RSF, remodeling and spacing factor; SNF2h, sucrose nonfermenting protein 2 homologue.

© 2009 Perpelescu et al.
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