DNA-protein cross-links (DPCs) are super-bulky, helix-distorting DNA lesions that result from exposure to a variety of chemical and physical agents such as antitumor drugs, environmental/ occupational toxins, ionizing radiation, and endogenous free radical-generating systems. Due to their unusually bulky nature, DPCs are hypothesized to interfere with normal DNA-protein interactions required for DNA replication, transcription, and repair, potentially leading to mutagenesis, genotoxicity, and cytotoxicity. However, the biological outcomes of DPC lesions in cells are poorly understood because of their inherent structural complexity and the difficulty of generating DNA substrates containing structurally defined DPC for biological evaluation. Our long-term goal is to elucidate the role tha spontaneous and induced DPCs play in human disease and in the anticancer activity of common chemotherapeutic agents. The objectives of this application are to determine the biological consequences of DPC formation in human cells and to discover the mechanism(s) responsible for the removal of DPCs from DNA. Our studies will for the first time systematically examine the influence of DPCs on DNA repair and replication. Our central hypothesis is that, if not repaired, DPCs exert mutagenic and cytotoxic effects, significantly contributing to the biological effects of many known carcinogens and antitumor drugs. Collectively, these studies will provide significant new insights into the molecular and cellular biology of DPCs and are expected to fundamentally advance the fields of DNA repair, cancer biology, and molecular mechanisms of aging.

Public Health Relevance

Exposure to certain anticancer drugs, environmental pollutants, and chemicals naturally produced in the body can cause proteins to become trapped on DNA. The resulting DNA-protein cross-links, if not repaired, can have toxic effects or lead to cancer, aging, and heart disease. The goals of this project are to understand how human cells cope with DNA-protein cross-links.

National Institute of Health (NIH)
National Institute of Environmental Health Sciences (NIEHS)
Research Project (R01)
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Special Emphasis Panel (ZRG1-OBT-S (02))
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Shaughnessy, Daniel
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University of Minnesota Twin Cities
Internal Medicine/Medicine
Schools of Medicine
United States
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Groehler 4th, Arnold; Villalta, Peter W; Campbell, Colin et al. (2016) Covalent DNA-Protein Cross-Linking by Phosphoramide Mustard and Nornitrogen Mustard in Human Cells. Chem Res Toxicol 29:190-202
Wickramaratne, Susith; Ji, Shaofei; Mukherjee, Shivam et al. (2016) Bypass of DNA-Protein Cross-links Conjugated to the 7-Deazaguanine Position of DNA by Translesion Synthesis Polymerases. J Biol Chem 291:23589-23603
Wickramaratne, Susith; Boldry, Emily J; Buehler, Charles et al. (2015) Error-prone translesion synthesis past DNA-peptide cross-links conjugated to the major groove of DNA via C5 of thymidine. J Biol Chem 290:775-87
Tretyakova, Natalia Y; Groehler 4th, Arnold; Ji, Shaofei (2015) DNA-Protein Cross-Links: Formation, Structural Identities, and Biological Outcomes. Acc Chem Res 48:1631-44
Wickramaratne, Susith; Seiler, Christopher L; Tretyakova, Natalia Y (2015) Synthesis of DNA Oligodeoxynucleotides Containing Site-Specific 1,3-Butadiene-Deoxyadenosine Lesions. Curr Protoc Nucleic Acid Chem 61:4.61.1-22
Weerasooriya, Savithri; Jasti, Vijay P; Basu, Ashis K (2014) Replicative bypass of abasic site in Escherichia coli and human cells: similarities and differences. PLoS One 9:e107915