Dynamic interaction of DNA damage checkpoint protein Rad53 with chromatin assembly factor Asf1.
The evolutionarily conserved yeast checkpoint protein kinase Rad53 regulates cell cycle progression, transcription, and DNA repair in response to DNA damage. To uncover potential regulatory targets of Rad53, we identified proteins physically associated with it in vivo using protein affinity purification and tandem mass spectrometry. Here we report that Rad53 interacts in a dynamic functional manner with Asf1, a chromatin assembly factor recently shown to mediate deposition of acetylated histones H3 and H4 onto newly replicated DNA. Biochemical and molecular genetic studies suggest that Asf1 is an important target of the Rad53-dependent DNA damage response and that Rad53 may directly regulate chromatin assembly during DNA replication and repair.
Emili
A
A
Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA. andrew.emili@utoronto.ca
Schieltz
D M
DM
Yates
J R
JR
Hartwell
L H
LH
D002453:Cell Cycle / Q000235:genetics; D018797:Cell Cycle Proteins / Q000378:metabolism*; D064447:Checkpoint Kinase 2; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology; D004271:DNA, Fungal / Q000502:physiology; D018816:Genes, cdc / Q000502:physiology*; D006657:Histones / Q000378:metabolism; D066298:In Vitro Techniques; D018832:Molecular Chaperones; D010766:Phosphorylation; D011485:Protein Binding / Q000235:genetics; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D029701:Saccharomyces cerevisiae Proteins; D015003:Yeasts
C109731:ASF1 protein, S cerevisiae; D018797:Cell Cycle Proteins; D002843:Chromatin; D004271:DNA, Fungal; D006657:Histones; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; C067022:RAD53 protein, S cerevisiae