Reactive oxygen species (ROS) can be induced by both endogenous and exogenous sources and they can result in DNA damage. We hypothesize that the ROS-induced bulky DNA lesions, owing to their difficulty in being repaired and their miscoding potential, may contribute significantly to the ubiquitously observed mutations at CpG dinucleotide site. To test this hypothesis, we will take an interdisciplinary approach, which encompasses synthetic organic chemistry, bioanalytical chemistry and molecular biology, to investigate the formation of these lesions in vitro and in vivo and to assess how they are interpreted by DNA replication and repair machineries in human cells. We organize the proposed research into three specific aims:
In Aim 1, we will assess quantitatively the formation of bulky lesions in DNA upon exposure to Fenton reagents and investigate how cytosine methylation affects the formation of 8,5'-cyclo-2'-deoxyguanosine at CpG site. We will also quantify these lesions formed in tissues of a rat model of Wilson's disease, which arises from excessive accumulation of copper ion in the liver.
In Aim 2, we will investigate the cytotoxic and mutagenic properties of these lesions in mammalian cells by using our newly developed shuttle vector technology.
In Aim 3, we will examine how the ROS-induced bulky DNA lesions are repaired in human cells. The significance of the proposed research lies in that it will provide important insights into the implications of these lesions in the development of human diseases including cancer and neurological disorders. Additionally, the results from the proposed research may reveal potential risk factors for developing human diseases and lead to the discovery of new molecular signatures for monitoring the human exposure toward ROS. The proposed research is highly innovative because it involves the use of a multi-pronged approach to tackle an important, yet largely overlooked biological problem.

Public Health Relevance

The focus of this proposal is to assess the formation and biological consequences bulky DNA lesions induced by reactive oxygen species (ROS). The outcome of the proposed research will improve significantly our understanding of the implications of ROS exposure in the development of cancer and neurological disorders and may lead to the discovery of novel biomarkers for monitoring the human exposure toward ROS.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA101864-09S1
Application #
8795263
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Lin, Alison J
Project Start
2003-07-01
Project End
2016-02-29
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
9
Fiscal Year
2014
Total Cost
$72,481
Indirect Cost
$24,796
Name
University of California Riverside
Department
Chemistry
Type
Schools of Earth Sciences/Natur
DUNS #
627797426
City
Riverside
State
CA
Country
United States
Zip Code
92521
Ji, Debin; Lin, Krystal; Song, Jikui et al. (2014) Effects of Tet-induced oxidation products of 5-methylcytosine on Dnmt1- and DNMT3a-mediated cytosine methylation. Mol Biosyst 10:1749-52
Fu, Lijuan; Guerrero, Candace R; Zhong, Na et al. (2014) Tet-mediated formation of 5-hydroxymethylcytosine in RNA. J Am Chem Soc 136:11582-5
Chavez, Lukas; Huang, Yun; Luong, Khai et al. (2014) Simultaneous sequencing of oxidized methylcytosines produced by TET/JBP dioxygenases in Coprinopsis cinerea. Proc Natl Acad Sci U S A 111:E5149-58
Wang, Pengcheng; Williams, Renee T; Guerrero, Candace R et al. (2014) Fragmentation of electrospray-produced deprotonated ions of oligodeoxyribonucleotides containing an alkylated or oxidized thymidine. J Am Soc Mass Spectrom 25:1167-76
Weldon, David J; Saulsbury, Marilyn D; Goh, Joshua et al. (2014) One-pot synthesis of cinnamylideneacetophenones and their in vitro cytotoxicity in breast cancer cells. Bioorg Med Chem Lett 24:3381-4
Amato, Nicholas J; Wang, Yinsheng (2014) Epimeric 2-deoxyribose lesions: Products from the improper chemical repair of 2-deoxyribose radicals. Chem Res Toxicol 27:470-9
Ji, Debin; You, Changjun; Wang, Pengcheng et al. (2014) Effects of tet-induced oxidation products of 5-methylcytosine on DNA replication in mammalian cells. Chem Res Toxicol 27:1304-9
You, Changjun; Ji, Debin; Dai, Xiaoxia et al. (2014) Effects of Tet-mediated oxidation products of 5-methylcytosine on DNA transcription in vitro and in mammalian cells. Sci Rep 4:7052
Ji, Debin; Wang, Yinsheng (2014) Facile enzymatic synthesis of base J-containing oligodeoxyribonucleotides and an analysis of the impact of base J on DNA replication in cells. PLoS One 9:e103335
Bullard, Whitney; Lopes da Rosa-Spiegler, Jessica; Liu, Shuo et al. (2014) Identification of the glucosyltransferase that converts hydroxymethyluracil to base J in the trypanosomatid genome. J Biol Chem 289:20273-82

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