Nucleic acid oxidation is important in the etiology and treatment of disease. For instance, ionizing radiation causes cancer and destroys tumor cells by damaging DNA. DNA damage is also involved in aging and a variety of other diseases (e.g. Xeroderma Pigmentosum, cystic fibrosis, myocardial infarction). The goals of this research are to understand how lesions produced in DNA as a result of oxidative stress are repaired, replicated, and react to form other lesions. We are also developing tools for selectively detecting DNA lesions. Much of our effort focuses on oxidized abasic lesions, which are incapable of forming Watson-Crick hydrogen bonds. Contrary to what was previously believed, oxidized abasic lesions interact with polymerases in distinct ways from each other and from an abasic site (AP) resulting from formal hydrolysis of a nucleotide's glycosidic bond. Hence, the inability to form Watson-Crick hydrogen bonds does not mean that a lesion is noninstructive. We will use synthetic chemistry to synthesize analogues, rapid-quench kinetics to determine polymerase mechanisms, as well as shuttle vector experiments to determine what structural properties of the individual lesions give rise to their observed effects in cells. We will also examine the repair of lesions that would appear to require unusual handling by DNA repair enzymes. Finally, we will develop and employ reagents that enable us to selectively detect oxidized abasic lesions. These tools will enable scientists to correlate unique biological effects of lesions with their formation by various oxidizing agents. Relevance to public health: Oxidative nucleic acid damage plays an important role in aging, as well as the etiology and treatment of genetic diseases, such as cancer. This fundamental research is valuable to understanding the etiology and treatment of diseases such as cancer. Furthermore, the reagents developed by us for selectively detecting oxidized abasic sites will be valuable biotechnology tools. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM063028-08
Application #
7485844
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Fabian, Miles
Project Start
2001-04-01
Project End
2010-08-31
Budget Start
2008-09-01
Budget End
2009-08-31
Support Year
8
Fiscal Year
2008
Total Cost
$324,199
Indirect Cost
Name
Johns Hopkins University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Rana, Anup; Yang, Kun; Greenberg, Marc M (2018) Reactivity of the Major Product of C5'-Oxidative DNA Damage in Nucleosome Core Particles. Chembiochem :
Bai, Jing; Zhang, Yingqian; Xi, Zhen et al. (2018) Oxidation of 8-Oxo-7,8-dihydro-2'-deoxyguanosine Leads to Substantial DNA-Histone Cross-Links within Nucleosome Core Particles. Chem Res Toxicol :
Laverty, Daniel J; Greenberg, Marc M (2018) Expanded Substrate Scope of DNA Polymerase ? and DNA Polymerase ?: Lyase Activity on 5'-Overhangs and Clustered Lesions. Biochemistry 57:6119-6127
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
Yang, Kun; Greenberg, Marc M (2018) Enhanced Cleavage at Abasic Sites within Clustered Lesions in Nucleosome Core Particles. Chembiochem 19:2061-2065
Beaver, Jill M; Lai, Yanhao; Rolle, Shantell J et al. (2018) An oxidized abasic lesion inhibits base excision repair leading to DNA strand breaks in a trinucleotide repeat tract. PLoS One 13:e0192148
Laverty, Daniel J; Mortimer, Ifor P; Greenberg, Marc M (2018) Mechanistic Insight through Irreversible Inhibition: DNA Polymerase ? Uses a Common Active Site for Polymerase and Lyase Activities. J Am Chem Soc 140:9034-9037
Jacinto, Marco Paolo; Pichling, Patricio; Greenberg, Marc M (2018) Synthesis of 5-Methylene-2-pyrrolones. Org Lett 20:4885-4887
Wang, Ruixiang; Yang, Kun; Banerjee, Samya et al. (2018) Rotational Effects within Nucleosome Core Particles on Abasic Site Reactivity. Biochemistry 57:3945-3952
Laverty, Daniel J; Averill, April M; DoubliƩ, Sylvie et al. (2017) The A-Rule and Deletion Formation During Abasic and Oxidized Abasic Site Bypass by DNA Polymerase ?. ACS Chem Biol 12:1584-1592

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