Abstract

Maintenance of genome stability is essential for cell survival and is critical for the prevention of carcinogenesis. Indeed, genetic instability is one of the key features of cancer cells. Because errors in DNA replication are a major cause of genomic instability, it is critical to understand all aspects involved in the regulation of DNA replication. However, relatively few studies have addressed how chromatin structure, the physiological organization of DNA in eukaryotic cells, impacts and regulates the DNA replication process. During DNA replication, chromatin must be temporarily disassembled to allow the DNA replication machinery access to the DNA template. The newly-synthesized DNA strands are then immediately reassembled into their original chromatin state. Uncoupling of DNA synthesis and chromatin assembly results in genome instability. Because of the critical nature of genome stability, it is important to understand how DNA synthesis is coupled to chromatin assembly. We use the yeast Saccharomyces cerevisiae as a eukaryotic model organism to address questions regarding this essential, but poorly understood process. Recent studies have implicated the acetylation of lysine 56 on histone H3 (H3-K56) as a key regulatory event during the DNA synthesis and chromatin assembly. Acetylation of H3-K56 occurs transiently during S phase. Cells with altered acetylation of H3-K56 are highly sensitive to DNA damaging agents, suggesting that this modification is important for maintaining genome stability. The main objective of this proposal is to test the hypothesis that acetylation of H3-K56 by a novel and unique HAT Rtt109, which we have recently identified, maintains genome integrity by coupling DNA replication with the assembly of newly-replicated DNA into nucleosomes mediated by histone chaperones. Results from these studies will provide mechanistic insights into the role of histone modifications in coupling DNA synthesis with nucleosome assembly, an important aspect in the maintenance of genome stability and thus normal cell functions.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM081838-03
Application #
7798234
Study Section
Molecular Genetics A Study Section (MGA)
Program Officer
Carter, Anthony D
Project Start
2008-05-01
Project End
2012-03-31
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
3
Fiscal Year
2010
Total Cost
$291,699
Indirect Cost

  Institution

Name
Mayo Clinic, Rochester
Department
Type
DUNS #
006471700
City
Rochester
State
MN
Country
United States
Zip Code
55905

  Publications

Zhang, Honglian; Gan, Haiyun; Wang, Zhiquan et al. (2017) RPA Interacts with HIRA and Regulates H3.3 Deposition at Gene Regulatory Elements in Mammalian Cells. Mol Cell 65:272-284
Fang, Dong; Gan, Haiyun; Wang, Heping et al. (2017) Probe the function of histone lysine 36 methylation using histone H3 lysine 36 to methionine mutant transgene in mammalian cells. Cell Cycle 16:1781-1789
Zhang, Kuo; Gao, Yuan; Li, Jingjing et al. (2016) A DNA binding winged helix domain in CAF-1 functions with PCNA to stabilize CAF-1 at replication forks. Nucleic Acids Res 44:5083-94
Feng, Jianxun; Gan, Haiyun; Eaton, Matthew L et al. (2016) Noncoding Transcription Is a Driving Force for Nucleosome Instability in spt16 Mutant Cells. Mol Cell Biol 36:1856-67
Wang, Zhiquan; Zhang, Honglian; Liu, Ji et al. (2016) USP51 deubiquitylates H2AK13,15ub and regulates DNA damage response. Genes Dev 30:946-59
Fang, Dong; Gan, Haiyun; Lee, Jeong-Heon et al. (2016) The histone H3.3K36M mutation reprograms the epigenome of chondroblastomas. Science 352:1344-8
Dahlin, Jayme L; Nissink, J Willem M; Strasser, Jessica M et al. (2015) PAINS in the assay: chemical mechanisms of assay interference and promiscuous enzymatic inhibition observed during a sulfhydryl-scavenging HTS. J Med Chem 58:2091-113
Dahlin, Jayme L; Nissink, J Willem M; Francis, Subhashree et al. (2015) Post-HTS case report and structural alert: Promiscuous 4-aroyl-1,5-disubstituted-3-hydroxy-2H-pyrrol-2-one actives verified by ALARM NMR. Bioorg Med Chem Lett 25:4740-4752
Dahlin, Jayme L; Chen, Xiaoyue; Walters, Michael A et al. (2015) Histone-modifying enzymes, histone modifications and histone chaperones in nucleosome assembly: Lessons learned from Rtt109 histone acetyltransferases. Crit Rev Biochem Mol Biol 50:31-53
Yu, Chuanhe; Gan, Haiyun; Han, Junhong et al. (2014) Strand-specific analysis shows protein binding at replication forks and PCNA unloading from lagging strands when forks stall. Mol Cell 56:551-63

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