Class I Histone Deacetylase HDAC1 and WRN RECQ Helicase Contribute Additively to Protect Replication Forks upon Hydroxyurea-induced Arrest.
The WRN helicase/exonuclease is mutated in Werner syndrome of genomic instability and premature aging. WRN-depleted fibroblasts, although remaining largely viable, have a reduced capacity to maintain replication forks active during a transient hydroxyurea-induced arrest. A strand exchange protein, RAD51, is also required for replication fork maintenance, and here we show that recruitment of RAD51 to stalled forks is reduced in the absence of WRN. We performed a siRNA screen for genes that are required for viability of WRN-depleted cells after hydroxyurea treatment, and identified HDAC1, a member of the class I histone deacetylase family. One of the functions of HDAC1, which it performs together with a close homolog HDAC2, is deacetylation of new histone H4 deposited at replication forks. We show that HDAC1 depletion exacerbates defects in fork reactivation and progression after hydroxyurea treatment observed in WRN- or RAD51-deficient cells. The additive WRN, HDAC1 loss-of-function phenotype is also observed with a catalytic mutant of HDAC1; however, it does not correlate with changes in histone H4 deacetylation at replication forks. On the other hand, inhibition of histone deacetylation by an inhibitor specific to HDACs 1-3, CI-994, correlates with increased processing of newly synthesized DNA strands in hydroxyurea-stalled forks. WRN co-precipitates with HDAC1 and HDAC2. Taken together, our findings indicate that WRN interacts with HDACs 1 and 2 to facilitate activity of stalled replication forks under conditions of replication stress.
From the Department of Pathology and.
Keffy
K
Kehrli
From the Department of Pathology and.
Michael
M
Phelps
From the Department of Pathology and.
Pavlo
P
Lazarchuk
From the Department of Pathology and.
Eleanor
E
Chen
From the Department of Pathology and; Department of Genome Sciences, University of Washington, Seattle, Washington 98195.
Ray
R
Monnat
From the Department of Pathology and. Electronic address: julias@uw.edu.
Julia M
JM
Sidorova
D059447:Cell Cycle Checkpoints / Q000187:drug effects* / Q000235:genetics; D002461:Cell Line, Transformed; D004261:DNA Replication / Q000187:drug effects* / Q000235:genetics; D056284:Histone Deacetylase 1 / Q000235:genetics / Q000378:metabolism*; D056464:Histone Deacetylase 2 / Q000235:genetics / Q000378:metabolism; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology*; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D000071657:Werner Syndrome Helicase / Q000235:genetics / Q000378:metabolism*; D000099186:Histone Deacetylase 3
D056284:Histone Deacetylase 1; D056464:Histone Deacetylase 2; D006655:Histone Deacetylases; D006918:Hydroxyurea; D051135:Rad51 Recombinase; D000071657:Werner Syndrome Helicase; D000099186:Histone Deacetylase 3; C495153:RAD51 protein, human; C110550:HDAC1 protein, human; C533423:HDAC2 protein, human; C098785:WRN protein, human
DNA helicase; DNA replication; Werner syndrome protein; histone acetylation; histone deacetylase 1 (HDAC1); human; small interfering RNA (siRNA); tissue culture
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