TY - JOUR
T1 - Histone deacetylases 1 and 2 maintain S-phase chromatin and DNA replication fork progression
AU - Bhaskara, Srividya
AU - Jacques, Vincent
AU - Rusche, James R.
AU - Olson, Eric N.
AU - Cairns, Bradley R.
AU - Chandrasekharan, Mahesh B.
N1 - Funding Information:
We thank David Jones and Maria McDowell for their comments on the manuscript. We thank Hannah Wellman, Jean Ansolabehere, Brandt Jones and Saravanan Ramakrishnan for their technical assistance. We thank Cedric Clapier and Takuya Abe for their suggestions. We thank David Nix and Brian Dalley at the HCI genomics core for assistance with the RNA-seq analysis. We are grateful to Kim Boucher for the statistical analyses. We thank Jun Qin and Beom-Jun Kim for the Smc3 cell lines and Katsuhiko Shirahige for the Smc3ac antibody. We specially thank Scott Hiebert for his kind encouragement and support of this work, and comments on this manuscript. This work was supported by the Huntsman Cancer Institute and Radiation Oncology Department Development Funds provided to SB.
PY - 2013
Y1 - 2013
N2 - Background: Histone deacetylases (HDACs) play a critical role in the maintenance of genome stability. Class I HDACs, histone deacetylase 1 and 2 (Hdac1 and Hdac2) are recruited to the replication fork by virtue of their interactions with the replication machinery. However, functions for Hdac1 and Hdac2 (Hdacs1,2) in DNA replication are not fully understood. Results: Using genetic knockdown systems and novel Hdacs1,2-selective inhibitors, we found that loss of Hdacs1,2 leads to a reduction in the replication fork velocity, and an increase in replication stress response culminating in DNA damage. These observed defects are due to a direct role for Hdacs1,2 in DNA replication, as transcription of genes involved in replication was not affected in the absence of Hdacs1,2. We found that loss of Hdacs1,2 functions increases histone acetylation (ac) on chromatin in S-phase cells and affects nascent chromatin structure, as evidenced by the altered sensitivity of newly synthesized DNA to nuclease digestion. Specifically, H4K16ac, a histone modification involved in chromatin decompaction, is increased on nascent chromatin upon abolishing Hdacs1,2 activities. It was previously shown that H4K16ac interferes with the functions of SMARCA5, an ATP-dependent ISWI family chromatin remodeler. We found SMARCA5 also associates with nascent DNA and loss of SMARCA5 decreases replication fork velocity similar to the loss or inhibition of Hdacs1,2. Conclusions: Our studies reveal important roles for Hdacs1,2 in nascent chromatin structure maintenance and regulation of SMARCA5 chromatin-remodeler function, which together are required for proper replication fork progression and genome stability in S-phase.
AB - Background: Histone deacetylases (HDACs) play a critical role in the maintenance of genome stability. Class I HDACs, histone deacetylase 1 and 2 (Hdac1 and Hdac2) are recruited to the replication fork by virtue of their interactions with the replication machinery. However, functions for Hdac1 and Hdac2 (Hdacs1,2) in DNA replication are not fully understood. Results: Using genetic knockdown systems and novel Hdacs1,2-selective inhibitors, we found that loss of Hdacs1,2 leads to a reduction in the replication fork velocity, and an increase in replication stress response culminating in DNA damage. These observed defects are due to a direct role for Hdacs1,2 in DNA replication, as transcription of genes involved in replication was not affected in the absence of Hdacs1,2. We found that loss of Hdacs1,2 functions increases histone acetylation (ac) on chromatin in S-phase cells and affects nascent chromatin structure, as evidenced by the altered sensitivity of newly synthesized DNA to nuclease digestion. Specifically, H4K16ac, a histone modification involved in chromatin decompaction, is increased on nascent chromatin upon abolishing Hdacs1,2 activities. It was previously shown that H4K16ac interferes with the functions of SMARCA5, an ATP-dependent ISWI family chromatin remodeler. We found SMARCA5 also associates with nascent DNA and loss of SMARCA5 decreases replication fork velocity similar to the loss or inhibition of Hdacs1,2. Conclusions: Our studies reveal important roles for Hdacs1,2 in nascent chromatin structure maintenance and regulation of SMARCA5 chromatin-remodeler function, which together are required for proper replication fork progression and genome stability in S-phase.
KW - Chromatin remodelers
KW - Hdacs1
KW - Hdacs2
KW - Nascent chromatin
KW - Replication
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U2 - 10.1186/1756-8935-6-27
DO - 10.1186/1756-8935-6-27
M3 - Article
C2 - 23947532
AN - SCOPUS:84881419689
SN - 1756-8935
VL - 6
JO - Epigenetics and Chromatin
JF - Epigenetics and Chromatin
IS - 1
M1 - 27
ER -