The RCAF complex mediates chromatin assembly during DNA replication and repair.
Chromatin assembly is a fundamental biological process that is essential for the replication and maintenance of the eukaryotic genome. In dividing cells, newly synthesized DNA is rapidly assembled into chromatin by the deposition of a tetramer of the histone proteins H3 and H4, followed by the deposition of two dimers of histones H2A and H2B to complete the nucleosome-the fundamental repeating unit of chromatin. Here we describe the identification, purification, cloning, and characterization of replication-coupling assembly factor (RCAF), a novel protein complex that facilitates the assembly of nucleosomes onto newly replicated DNA in vitro. RCAF comprises the Drosophila homologue of anti-silencing function 1 protein ASF1 and histones H3 and H4. The specific acetylation pattern of H3 and H4 in RCAF is identical to that of newly synthesized histones. Genetic analyses in Saccharomyces cerevisiae demonstrate that ASF1 is essential for normal cell cycle progression, and suggest that RCAF mediates chromatin assembly after DNA replication and the repair of double-strand DNA damage in vivo.
Tyler
J K
JK
Department of Biology and Center for Molecular Genetics, University of California at San Diego, La Jolla 92093-0347, USA.
Adams
C R
CR
Chen
S R
SR
Kobayashi
R
R
Kamakaka
R T
RT
Kadonaga
J T
JT
D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D002843:Chromatin / Q000378:metabolism*; D003001:Cloning, Molecular; D004260:DNA Repair; D004261:DNA Replication; D004330:Drosophila / Q000235:genetics / Q000502:physiology*; D029721:Drosophila Proteins; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D018832:Molecular Chaperones / Q000502:physiology; D008969:Molecular Sequence Data
C491641:ASF1A protein, human; C491640:ASF1B protein, human; D018797:Cell Cycle Proteins; D002843:Chromatin; D029721:Drosophila Proteins; D018832:Molecular Chaperones; C491642:asf1 protein, Drosophila