Malondialdehyde (MDA) and other aldehydic products of oxidative stress represent an important class of endogenous DMA damaging agents universally generated in human beings. These bifunctional electrophiles generate exocyclic adducts to deoxynucleoside bases that prevent Watson-Crick base pairing, block DNA replication, and induce mutations. Our laboratory has defined the chemistry and biology of the pyrimidopurinone adduct to deoxyguanosine, M1dG, which is the major product of DNA damage by MDA and base propenals. Duplex DNA catalyzes a dynamic equilibrium between M1dG and its ring-opened derivative, N2-(3-oxopropenyl)-deoxyguanosine (OPdG). OPdG is less blocking than M1dG to DNA replication and less mutagenic than stable exocyclic analogs of M1dG . We recently discovered that M1dG induces sequence-dependent frameshift mutations in reiterated d(CG)n sequences in E. coli and mammalian cells. We hypothesize that the ability of M1dG to induce frameshift mutations is due to its conversion to OPdG and we propose to test this hypothesis by directly comparing the ability of a series of structural analogs of M1dG and OPdG to induce frameshift mutations in d(CG)n sequences. We recently developed a general synthesis of MDA-DNA adducts in olignucleotides and we propose to use it to explore the chemistry and biology of the other major DNA adduct, N6-(3-oxopropenyl)-deoxyadenosine (OPdA). We will determine the mutation spectrum of OPdA in mammalian cells and we will test the hypothesis that OPdA and other deoxyadenosine products of oxidative stress induce frameshift mutations in reiterated dAn sequences. This type of mutation is commonly observed in critical growth regulating genes that contribute to human cancers and its occurrence has been associated with oxidative stress. MDA-induced DNA-protein cross-links have been reported to exist in cells exposed to oxidants but the identity of these cross-links and the chemistry of their formation is very poorly defined. We propose to explore the chemistry of reaction of M1dG, OPdG, and OPdA with amino acids, peptides, and proteins. Particular attention will be given to the identification of conjugates with DMA-binding proteins such as restriction endonucleases, nucleotide excision repair enzymes, and histones. These experiments will provide critical chemical information with which to evaluate the hypothesis that MDA-induced DNA-protein cross-links are important products of oxidative damage to cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
2R37CA087819-06
Application #
6926428
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Okano, Paul
Project Start
2000-06-01
Project End
2010-05-31
Budget Start
2005-06-01
Budget End
2006-05-31
Support Year
6
Fiscal Year
2005
Total Cost
$414,034
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Biochemistry
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Galligan, James J; Wepy, James A; Streeter, Matthew D et al. (2018) Methylglyoxal-derived posttranslational arginine modifications are abundant histone marks. Proc Natl Acad Sci U S A 115:9228-9233
Galligan, James J; Marnett, Lawrence J (2017) Histone Adduction and Its Functional Impact on Epigenetics. Chem Res Toxicol 30:376-387
Galligan, James J; Kingsley, Philip J; Wauchope, Orrette R et al. (2017) Quantitative Analysis and Discovery of Lysine and Arginine Modifications. Anal Chem 89:1299-1306
Beavers, William N; Rose, Kristie L; Galligan, James J et al. (2017) Protein Modification by Endogenously Generated Lipid Electrophiles: Mitochondria as the Source and Target. ACS Chem Biol 12:2062-2069
Camarillo, Jeannie M; Ullery, Jody C; Rose, Kristie L et al. (2017) Electrophilic Modification of PKM2 by 4-Hydroxynonenal and 4-Oxononenal Results in Protein Cross-Linking and Kinase Inhibition. Chem Res Toxicol 30:635-641
Camarillo, Jeannie M; Rose, Kristie L; Galligan, James J et al. (2016) Covalent Modification of CDK2 by 4-Hydroxynonenal as a Mechanism of Inhibition of Cell Cycle Progression. Chem Res Toxicol 29:323-32
Wauchope, Orrette R; Beavers, William N; Galligan, James J et al. (2015) Nuclear Oxidation of a Major Peroxidation DNA Adduct, M1dG, in the Genome. Chem Res Toxicol 28:2334-42
Beavers, William N; Serwa, Remigiusz; Shimozu, Yuki et al. (2014) ?-Alkynyl lipid surrogates for polyunsaturated fatty acids: free radical and enzymatic oxidations. J Am Chem Soc 136:11529-39
Galligan, James J; Rose, Kristie L; Beavers, William N et al. (2014) Stable histone adduction by 4-oxo-2-nonenal: a potential link between oxidative stress and epigenetics. J Am Chem Soc 136:11864-6
Shuck, Sarah C; Rose, Kristie L; Marnett, Lawrence J (2014) Mass spectrometric methods for the analysis of nucleoside-protein cross-links: application to oxopropenyl-deoxyadenosine. Chem Res Toxicol 27:136-46

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