The long-term goal of the proposed research is to understand how cells preserve genome integrity during DNA replication. Specifically, this application focuses on the ATR (ATM and rad3-related) signaling pathway. ATR functions at the apex of a DNA damage and replication stress response pathway that is needed every cell division cycle to promote the complete and accurate replication of the genome. Many cancer cells are highly dependent on ATR function for proliferation and viability because of elevated levels of oncogene-induced replication stress and mutations in other genome maintenance pathways. Thus, ATR may be a useful drug target based on a synthetic lethal approach. In this proposal we test a specific model of how ATR promotes replication fork stabilization and repair, define the consequences of acute ATR inhibition on DNA replication and cell fate outcomes, define and characterize new ATR-regulated proteins that act at damaged replication forks, characterize cancer settings in which ATR inhibition might be useful using synthetic lethality, and test a new model of how ATR is regulated by autophosphorylation. This is a focused proposal aimed at understanding the most important and least understood aspects of ATR function. Specific hypotheses and innovative concepts based on preliminary data are tested using advanced biochemical and genetic approaches. In addition, the aims provide opportunities for unexpected discoveries about the mechanisms that maintain the genome during DNA replication and when ATR pathway inhibitors may be useful in the cancer clinic.

Public Health Relevance

The replication stress response controlled by the ATR kinase is essential to maintain genome integrity and prevent carcinogenesis. ATR pathway inhibitors are currently being developed as anti-cancer agents. This research proposal will define mechanisms by which ATR promotes genome maintenance during DNA replication and identify cellular consequences of acute disruption of this pathway.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA102729-11A1
Application #
8628242
Study Section
Cellular Signaling and Regulatory Systems Study Section (CSRS)
Program Officer
Pelroy, Richard
Project Start
2003-07-01
Project End
2019-04-30
Budget Start
2014-05-06
Budget End
2015-04-30
Support Year
11
Fiscal Year
2014
Total Cost
$264,458
Indirect Cost
$95,834
Name
Vanderbilt University Medical Center
Department
Biochemistry
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Badu-Nkansah, Akosua; Mason, Aaron C; Eichman, Brandt F et al. (2016) Identification of a Substrate Recognition Domain in the Replication Stress Response Protein Zinc Finger Ran-binding Domain-containing Protein 3 (ZRANB3). J Biol Chem 291:8251-7
Dungrawala, Huzefa; Rose, Kristie L; Bhat, Kamakoti P et al. (2015) The Replication Checkpoint Prevents Two Types of Fork Collapse without Regulating Replisome Stability. Mol Cell 59:998-1010
Carroll, Clinton; Hunley, Tracy E; Guo, Yan et al. (2015) A novel splice site mutation in SMARCAL1 results in aberrant exon definition in a child with Schimke immunoosseous dysplasia. Am J Med Genet A 167A:2260-4
Kavanaugh, Gina; Ye, Fei; Mohni, Kareem N et al. (2015) A whole genome RNAi screen identifies replication stress response genes. DNA Repair (Amst) 35:55-62
Kavanaugh, Gina; Zhao, Runxiang; Guo, Yan et al. (2015) Enhancer of Rudimentary Homolog Affects the Replication Stress Response through Regulation of RNA Processing. Mol Cell Biol 35:2979-90
Mohni, Kareem N; Thompson, Petria S; Luzwick, Jessica W et al. (2015) A Synthetic Lethal Screen Identifies DNA Repair Pathways that Sensitize Cancer Cells to Combined ATR Inhibition and Cisplatin Treatments. PLoS One 10:e0125482
Cortez, David (2015) Preventing replication fork collapse to maintain genome integrity. DNA Repair (Amst) 32:149-57
Sowd, Gregory A; Mody, Dviti; Eggold, Joshua et al. (2014) SV40 utilizes ATM kinase activity to prevent non-homologous end joining of broken viral DNA replication products. PLoS Pathog 10:e1004536
Frank, Andreas O; Vangamudi, Bhavatarini; Feldkamp, Michael D et al. (2014) Discovery of a potent stapled helix peptide that binds to the 70N domain of replication protein A. J Med Chem 57:2455-61
Mohni, Kareem N; Kavanaugh, Gina M; Cortez, David (2014) ATR pathway inhibition is synthetically lethal in cancer cells with ERCC1 deficiency. Cancer Res 74:2835-45

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