Activation of the DNA damage checkpoint in yeast lacking the histone chaperone anti-silencing function 1.
The packaging of the eukaryotic genome into chromatin is likely to be important for the maintenance of genomic integrity. Chromatin structures are assembled onto newly synthesized DNA by the action of chromatin assembly factors, including anti-silencing function 1 (ASF1). To investigate the role of chromatin structure in the maintenance of genomic integrity, we examined budding yeast lacking the histone chaperone Asf1p. We found that yeast lacking Asf1p accumulate in metaphase of the cell cycle due to activation of the DNA damage checkpoint. Furthermore, yeast lacking Asf1p are highly sensitive to mutations in DNA polymerase alpha and to DNA replicational stresses. Although yeast lacking Asf1p do complete DNA replication, they have greatly elevated rates of DNA damage occurring during DNA replication, as indicated by spontaneous Ddc2p-green fluorescent protein foci. The presence of elevated levels of spontaneous DNA damage in asf1 mutants is due to increased DNA damage, rather than the failure to repair double-strand DNA breaks, because asf1 mutants are fully functional for double-strand DNA repair. Our data indicate that the altered chromatin structure in asf1 mutants leads to elevated rates of spontaneous recombination, mutation, and DNA damage foci formation arising during DNA replication, which in turn activates cell cycle checkpoints that respond to DNA damage.
Ramey
Christopher Josh
CJ
Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center at Fitzsimons, P.O. Box 6511, Aurora, CO 80045, USA.
Howar
Susan
S
Adkins
Melissa
M
Linger
Jeffrey
J
Spicer
Judson
J
Tyler
Jessica K
JK
D048868:Adaptor Proteins, Signal Transducing; D015153:Blotting, Western; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002455:Cell Division; D049109:Cell Proliferation; D002470:Cell Survival; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D005434:Flow Cytometry; D016195:G2 Phase; D017353:Gene Deletion; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006657:Histones / Q000737:chemistry*; D008677:Metaphase; D008957:Models, Genetic; D018832:Molecular Chaperones; D009154:Mutation; D010750:Phosphoproteins / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics; D013696:Temperature; D013997:Time Factors
C109731:ASF1 protein, S cerevisiae; C491641:ASF1A protein, human; D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D002843:Chromatin; D006657:Histones; C440014:LCD1 protein, S cerevisiae; D018832:Molecular Chaperones; D010750:Phosphoproteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D004259:DNA-Directed DNA Polymerase