Active establishment of centromeric CENP-A chromatin by RSF complex.
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.
Perpelescu
Marinela
M
Bioscience and Biotechnology Center, Nagoya University, Nagoya, Aichi, Japan.
Nozaki
Naohito
N
Obuse
Chikashi
C
Yang
Hua
H
Yoda
Kinya
K
D001324:Autoantigens / Q000302:isolation & purification / Q000502:physiology*; D002503:Centromere / Q000235:genetics / Q000502:physiology; D000076247:Centromere Protein A; D002843:Chromatin / Q000235:genetics / Q000502:physiology*; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification / Q000502:physiology*; D004261:DNA Replication; D016193:G1 Phase; D006367:HeLa Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D007399:Interphase; D008938:Mitosis; D009687:Nuclear Proteins / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D015534:Trans-Activators / Q000502:physiology*; D014158:Transcription, Genetic
D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; C452851:RSF1 protein, human; D015534:Trans-Activators