The broad long-term goal of this project is to elucidate the molecular basis of environmental Polycyclic Aryl-Hydrocarbon (PAH)-induced cell cycle checkpoint control. We have identified a novel cell cycle checkpoint resulting from aryl-hydrocarbon-induced DNA damage. The PAH- activated checkpoint arrests proliferating cells in the G1 phase of the cell cycle. Growth arrest prior to S-phase prevents potentially error- prone and mutagenic replication of damaged DNA. Experiments proposed here seek to understand the molecular mechanism(s) whereby the PAH- induced checkpoint is imposed upon cells and, ultimately, how this mechanism is escaped during transformation. Our preliminary studies suggested new and important roles for tumor suppressors (Retinoblastoma, or Rb, and p53) and G1 signal transduction pathways in PAH-induced checkpoint control. The Rb and p53 genes play important roles in guarding against malignancy and are mutated or absent in many human cancers. Therefore, our preliminary data provided a new and direct link between the cellular effects of environmental PAHs and human malignancies. These studies will test the putative roles of Rb, G1 mitogenic signal transduction events, and p53 in regulating cellular responses to PAHs.
The specific aims of this project are: (1) To test the hypothesis that the PAH-induced cell cycle checkpoint is Rb- mediated. (2) To test the hypothesis that the PAH-induced checkpoint results from modification of mitogenic G1 signal transduction events. (3) To test the hypothesis that p53 plays a role in repair of PAH- adducted DNA. We will use loss-of-function (expression of viral oncogenes) and gain-of-function (ectopic expression of tumor suppressors in knockout tumor suppressor-deficient cells) strategies to test the roles of p53 and Rb in cellular responses to PAHs. Additionally, we will analyze the PAH-sensitivity of mitogenic signaling cascades during G1 in order to identify the putative PAH-induced lesion(s). These studies will identify novel molecular mechanisms of checkpoint control. This information may enable the rational design of new chemotherapies to manipulate checkpoint pathways. Such drugs could help prevent and treat PAH-induced (and possibly other) malignancies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29ES009558-04
Application #
6382264
Study Section
Chemical Pathology Study Section (CPA)
Program Officer
Packenham, Joan P
Project Start
1998-08-01
Project End
2002-07-31
Budget Start
2001-08-01
Budget End
2002-07-31
Support Year
4
Fiscal Year
2001
Total Cost
$110,514
Indirect Cost
Name
Boston University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Yang, Yang; Durando, Michael; Smith-Roe, Stephanie L et al. (2013) Cell cycle stage-specific roles of Rad18 in tolerance and repair of oxidative DNA damage. Nucleic Acids Res 41:2296-312
Barkley, Laura R; Palle, Komaraiah; Durando, Michael et al. (2012) c-Jun N-terminal kinase-mediated Rad18 phosphorylation facilitates Pol? recruitment to stalled replication forks. Mol Biol Cell 23:1943-54
Williams, Stacy A; Longerich, Simonne; Sung, Patrick et al. (2011) The E3 ubiquitin ligase RAD18 regulates ubiquitylation and chromatin loading of FANCD2 and FANCI. Blood 117:5078-87
Palle, Komaraiah; Vaziri, Cyrus (2011) Rad18 E3 ubiquitin ligase activity mediates Fanconi anemia pathway activation and cell survival following DNA Topoisomerase 1 inhibition. Cell Cycle 10:1625-38
Day, Tovah A; Palle, Komariah; Barkley, Laura R et al. (2010) Phosphorylated Rad18 directs DNA polymerase ? to sites of stalled replication. J Cell Biol 191:953-66
Song, Ihn Young; Barkley, Laura R; Day, Tovah A et al. (2010) A novel role for Fanconi anemia (FA) pathway effector protein FANCD2 in cell cycle progression of untransformed primary human cells. Cell Cycle 9:2375-88
Song, Ihn Young; Palle, Komaraiah; Gurkar, Aditi et al. (2010) Rad18-mediated translesion synthesis of bulky DNA adducts is coupled to activation of the Fanconi anemia DNA repair pathway. J Biol Chem 285:31525-36
Ohashi, Eiji; Hanafusa, Tomo; Kamei, Keijiro et al. (2009) Identification of a novel REV1-interacting motif necessary for DNA polymerase kappa function. Genes Cells 14:101-11
Barkley, Laura R; Song, Ihn Young; Zou, Ying et al. (2009) Reduced expression of GINS complex members induces hallmarks of pre-malignancy in primary untransformed human cells. Cell Cycle 8:1577-88
Ohmori, Haruo; Hanafusa, Tomo; Ohashi, Eiji et al. (2009) Separate roles of structured and unstructured regions of Y-family DNA polymerases. Adv Protein Chem Struct Biol 78:99-146

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