Human Asf1 regulates the flow of S phase histones during replicational stress.
Maintenance of chromosomal integrity requires tight coordination of histone biosynthesis with DNA replication. Here, we show that extracts from human cells exposed to replication stress display an increased capacity to support replication-coupled chromatin assembly. While in unperturbed S phase, hAsf1 existed in equilibrium between an active form and an inactive histone-free pool, replication stress mobilized the majority of hAsf1 into an active multichaperone complex together with histones. This active multichaperone complex was limiting for chromatin assembly in S phase extracts, and hAsf1 was required for the enhanced assembly activity in cells exposed to replication stress. Consistently, siRNA-mediated knockdown of hAsf1 impaired the kinetics of S phase progression. Together, these data suggest that hAsf1 provides the cells with a buffering system for histone excess generated in response to stalled replication and explains how mammalian cells maintain a critical "active" histone pool available for deposition during recovery from replication stresses.
Groth
Anja
A
Institute of Cancer Biology, The Danish Cancer Society, Strandboulevarden 49, DK-2100 Copenhagen, Denmark.
Ray-Gallet
Dominique
D
Quivy
Jean-Pierre
JP
Lukas
Jiri
J
Bartek
Jiri
J
Almouzni
Geneviève
G
D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002458:Cell Fractionation; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism; D004261:DNA Replication; D006657:Histones / Q000378:metabolism*; D006801:Humans; D006918:Hydroxyurea / Q000378:metabolism; D046911:Macromolecular Substances; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D019384:Nucleic Acid Synthesis Inhibitors / Q000378:metabolism; D016196:S Phase / Q000502:physiology*
C491641:ASF1A protein, human; D018797:Cell Cycle Proteins; D002843:Chromatin; D006657:Histones; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D019384:Nucleic Acid Synthesis Inhibitors; D006918:Hydroxyurea