DNA-protein cross-links (DPCs) are formed when proteins become covalently bound to DNA form spontaneously as a result of normal cellular processes such as lipid peroxidation, histone demethylation, DNA replication, transcription, and DNA repair. DPCs can be induced by exposure to anti-tumor drugs, transition metals, UV light, and ?-radiation. DPCs interfere with many biological processes and are implicated in the accelerated aging and increased cancer incidence observed in Ruijs-Aalfs syndrome patients. The goal of this application is to map DPC lesions along the genome, investigate how human cells recognize and remove these exceedingly bulky DPC lesions, and to identify the mechanisms by which they cause mutagenicity and cell death. Our central hypothesis is that unrepaired DPCs compromise the efficiency and accuracy of DNA replication and contribute to the toxicity and mutagenicity induced by the agents listed above. Our research plan focuses on three aims. First, will use next generation sequencing in combination with affinity pull down and protein precipitation to identify specific genomic regions susceptible to spontaneous and xenobiotic- induced DPC formation in human cells. Second, we will elucidate the role of proteolytic processing in DPC repair. Affinity capture, unbiased searches, and candidate gene-based approaches twill be used to identify proteins required for proteolytic processing and repair of DPCs, determine how cells convert DPCs to smaller peptide lesions (DpCs), and identity critical DNA repair proteins required for DPC removal from the genome. Third, we will investigate the effects of DPCs and DpCs on DNA replication. Our in vitro studies using DNA Pol ? showed that efficiency and fidelity of translesion synthesis past peptide DpCs is strongly dependent on DNA sequence context. We will examine the effects of sequence context on bypass efficiency and mutagenicity in human cells. The structural basis for the context effects on the efficiency and fidelity of bypass will be studied by molecular modeling and NMR studies. We will use a newly developed assay that employs pigyBac transposition of DpC or DPC containing DNA to examine the effects these lesions have on chromosome replication. These studies will for the first time examine the biological outcomes of structurally defined chromosomal DPCs in human cells.

Public Health Relevance

Exposure to a wide variety of agents including occupational and environmental pollutants, anticancer drugs, and chemical species produced naturally as a result of cellular metabolism can cause proteins to become covalently attached to DNA. Failure to repair these DNA-protein cross-links can result in adverse outcomes, including cell death, oncogenesis, and other age-related pathologies. The goals of this project are to understand how DNA protein crosslinks are recognized and repaired, as well as to understand mechanisms of damage tolerance.

National Institute of Health (NIH)
National Institute of Environmental Health Sciences (NIEHS)
Research Project (R01)
Project #
Application #
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Shaughnessy, Daniel
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Minnesota Twin Cities
Schools of Pharmacy
United States
Zip Code
Chesner, Lisa N; Campbell, Colin (2018) A quantitative PCR-based assay reveals that nucleotide excision repair plays a predominant role in the removal of DNA-protein crosslinks from plasmids transfected into mammalian cells. DNA Repair (Amst) 62:18-27
Groehler 4th, Arnold; Kren, Stefan; Li, Qinglu et al. (2018) Oxidative cross-linking of proteins to DNA following ischemia-reperfusion injury. Free Radic Biol Med 120:89-101
Basu, Ashis K (2018) DNA Damage, Mutagenesis and Cancer. Int J Mol Sci 19:
Yang, Kun; Park, Daeyoon; Tretyakova, Natalia Y et al. (2018) Histone tails decrease N7-methyl-2'-deoxyguanosine depurination and yield DNA-protein cross-links in nucleosome core particles and cells. Proc Natl Acad Sci U S A 115:E11212-E11220
Pujari, Suresh S; Zhang, Yi; Ji, Shaofei et al. (2018) Site-specific cross-linking of proteins to DNA via a new bioorthogonal approach employing oxime ligation. Chem Commun (Camb) 54:6296-6299
Mosa, Islam M; Pattammattel, Ajith; Kadimisetty, Karteek et al. (2017) Ultrathin Graphene-Protein Supercapacitors for Miniaturized Bioelectronics. Adv Energy Mater 7:
Basu, Ashis K; Pande, Paritosh; Bose, Arindam (2017) Translesion Synthesis of 2'-Deoxyguanosine Lesions by Eukaryotic DNA Polymerases. Chem Res Toxicol 30:61-72
Zore, Omkar V; Pande, Paritosh; Okifo, Oghenenyerovwo et al. (2017) Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis. RSC Adv 7:29563-29574
Groehler 4th, Arnold; Degner, Amanda; Tretyakova, Natalia Y (2017) Mass Spectrometry-Based Tools to Characterize DNA-Protein Cross-Linking by Bis-Electrophiles. Basic Clin Pharmacol Toxicol 121 Suppl 3:63-77
Pattammattel, Ajith; Pande, Paritosh; Kuttappan, Deepa et al. (2017) Controlling the Graphene-Bio Interface: Dispersions in Animal Sera for Enhanced Stability and Reduced Toxicity. Langmuir 33:14184-14194

Showing the most recent 10 out of 22 publications