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 #
3R01CA101864-08S1
Application #
8617321
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Ogunbiyi, Peter
Project Start
2003-07-01
Project End
2016-02-29
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
8
Fiscal Year
2013
Total Cost
$54,926
Indirect Cost
$14,258
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
You, Changjun; Wang, Yinsheng (2016) Mass Spectrometry-Based Quantitative Strategies for Assessing the Biological Consequences and Repair of DNA Adducts. Acc Chem Res 49:205-13
Yu, Yang; Guerrero, Candace R; Liu, Shuo et al. (2016) Comprehensive Assessment of Oxidatively Induced Modifications of DNA in a Rat Model of Human Wilson's Disease. Mol Cell Proteomics 15:810-7
Amato, Nicholas J; Zhai, Qianqian; Navarro, Diana C et al. (2015) In vivo detection and replication studies of α-anomeric lesions of 2'-deoxyribonucleosides. Nucleic Acids Res 43:8314-24
Zhang, Zhi-Min; Liu, Shuo; Lin, Krystal et al. (2015) Crystal Structure of Human DNA Methyltransferase 1. J Mol Biol 427:2520-31
Liu, Shuo; Wang, Yinsheng (2015) Mass spectrometry for the assessment of the occurrence and biological consequences of DNA adducts. Chem Soc Rev 44:7829-54
Fu, Lijuan; Amato, Nicholas J; Wang, Pengcheng et al. (2015) Simultaneous Quantification of Methylated Cytidine and Adenosine in Cellular and Tissue RNA by Nano-Flow Liquid Chromatography-Tandem Mass Spectrometry Coupled with the Stable Isotope-Dilution Method. Anal Chem 87:7653-9
McLean, Lancelot S; Watkins, Cheri N; Campbell, Petreena et al. (2015) Aryl Hydrocarbon Receptor Ligand 5F 203 Induces Oxidative Stress That Triggers DNA Damage in Human Breast Cancer Cells. Chem Res Toxicol 28:855-71
Wang, Xi-liang; Song, Shu-hui; Wu, Yong-Sheng et al. (2015) Genome-wide mapping of 5-hydroxymethylcytosine in three rice cultivars reveals its preferential localization in transcriptionally silent transposable element genes. J Exp Bot 66:6651-63
You, Changjun; Wang, Yinsheng (2015) Quantitative measurement of transcriptional inhibition and mutagenesis induced by site-specifically incorporated DNA lesions in vitro and in vivo. Nat Protoc 10:1389-406
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

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