Individuals who suffer from the genetic disease xeroderma pigmentosum (XP) lack nucleotide excision repair (NER) of DNA, and thus have much higher carcinogenic probability than the average population after exposure to UV irradiation. NER is a major cellular biological defense system to remove DNA damage due to the formation of bulky lesions induced by UV irradiation and environmental genotoxic chemicals and carcinogens. Although, it has been generally accepted that DNA damage recognition plays a central role in NER, the molecular and thermodynamic details of DNA damage processing remain largely unclear, and have not been systematically studied using biochemically rigorous approaches. In addition, the roles of the damage recognition proteins XPA, RPA, and XPC-HR23B have been controversial. The long-term objective of this study is to understand the molecular and biochemical details of UV-induced or related DNA damage recognition and repair by human NER proteins, and the potential effects of these relationships on damage-induced mutagenesis and carcinogenesis. To gain a systematic and more precise view of DNA damage recognition and repair by human NER, the following questions will be addressed: What is the hierarchy of damage recognition and how is damage dynamically processed in a stepwise recognition mechanism? What structural and chemical alterations in the DNA helix are identified by repair proteins at specific recognition steps? What is the molecular architecture of recognition intermediates? And what protein domains are important for protein-DNA and protein-protein contacts in the recognition. Specifically, this project aims to determine the thermodynamics and kinetics of the interaction of XPC-HR23B, XPA, and RPA with DNA substrates containing site-specific UV photolesions and benzo[a]pyrene diol epoxide (BPDE) DNA adducts using rigorous biochemical approaches: identify and analyze the important protein motifs involved in damage recognition: characterize the repair intermediates for damage recognition of UV-induced photolesions and BPDE-DNA adducts; and determine the mechanism in which the structural and chemical modifications of damage are recognized and repaired.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA086927-03
Application #
6514610
Study Section
Radiation Study Section (RAD)
Program Officer
Pelroy, Richard
Project Start
2000-07-01
Project End
2005-06-30
Budget Start
2002-07-01
Budget End
2003-06-30
Support Year
3
Fiscal Year
2002
Total Cost
$249,000
Indirect Cost
Name
East Tennessee State University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Johnson City
State
TN
Country
United States
Zip Code
37614
Wu, Xi; Dong, Zizheng; Wang, Chao J et al. (2016) FASN regulates cellular response to genotoxic treatments by increasing PARP-1 expression and DNA repair activity via NF-?B and SP1. Proc Natl Acad Sci U S A 113:E6965-E6973
Hilton, Benjamin; Gopal, Sathyaraj; Xu, Lifang et al. (2016) Dissociation Dynamics of XPC-RAD23B from Damaged DNA Is a Determining Factor of NER Efficiency. PLoS One 11:e0157784
Wang, Yan; Musich, Phillip R; Cui, Kui et al. (2015) Neurotoxin-induced DNA damage is persistent in SH-SY5Y cells and LC neurons. Neurotox Res 27:368-83
Hilton, Benjamin A; Li, Zhengke; Musich, Phillip R et al. (2015) ATR Plays a Direct Antiapoptotic Role at Mitochondria, which Is Regulated by Prolyl Isomerase Pin1. Mol Cell 60:35-46
Hilton, Benjamin; Shkriabai, Nick; Musich, Phillip R et al. (2014) A new structural insight into XPA-DNA interactions. Biosci Rep 34:e00162
Wang, Yan; Musich, Phillip R; Serrano, Moises A et al. (2014) Effects of DSP4 on the noradrenergic phenotypes and its potential molecular mechanisms in SH-SY5Y cells. Neurotox Res 25:193-207
Serrano, M A; Li, Z; Dangeti, M et al. (2013) DNA-PK, ATM and ATR collaboratively regulate p53-RPA interaction to facilitate homologous recombination DNA repair. Oncogene 32:2452-62
Jain, Vipin; Hilton, Benjamin; Lin, Bin et al. (2013) Structural and thermodynamic insight into Escherichia coli UvrABC-mediated incision of cluster diacetylaminofluorene adducts on the NarI sequence. Chem Res Toxicol 26:1251-62
Jain, Vipin; Hilton, Benjamin; Lin, Bin et al. (2013) Unusual sequence effects on nucleotide excision repair of arylamine lesions: DNA bending/distortion as a primary recognition factor. Nucleic Acids Res 41:869-80
Li, Zhengke; Musich, Phillip R; Cartwright, Brian M et al. (2013) UV-induced nuclear import of XPA is mediated by importin-?4 in an ATR-dependent manner. PLoS One 8:e68297

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