Mapping polycyclic aromatic hydrocarbon and aromatic amine-induced DNA damage in cancer-related genes at the sequence level

Authors: Tang M-s.1; Pfeifer G.P.2; Denissenko M.F.2; Feng Z.1; Hu W.1; Pao A.3; Zheng Y.3; Zheng J.B.3; Li H.3; Chen J.X.3

Source: International Journal of Hygiene and Environmental Health, Volume 205, Numbers 1-2, March 2002 , pp. 103-113(11)

Publisher: Urban & Fischer

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Abstract:

Genomic injury induced by environmental carcinogens, such as polycyclic aromatic hydrocarbons and aromatic amines, is the initial step that can trigger mutagenesis and carcinogenesis. In addition to the physico-chemical property of DNA damaging agents, several important factors such as primary sequence, chromatin structure, methylation, protein association, and transcriptional activity can affect not only the initial level and distribution of DNA damage but also the efficiency of repair. Therefore, mapping the DNA damage induced by environmental agents in cancer-related genes such as p53 and ras at the sequence level provides essential information for assessing their carcinogenic potential. Recently, using the E. coli nucleotide excision enzyme complex, UvrABC nucleases in combination with ligation-mediated polymerase chain reaction, we developed a method to map DNA damage in the p53 and ras genes. These studies led us to conclude that targeted DNA damage, in combination with growth selection, contributes greatly in shaping the mutation spectrum in these genes in human cancer. Here we present the rationale and details of this approach, typical experimental results and necessary precautions.

Keywords: Sequencing; DNA; damage; P53; ras

Language: English

Document Type: Original article

DOI: 10.1078/1438-4639-00135

Affiliations: 1: Department of Environmental Medicine, New York University School of Medicine, Tuxedo, New York 10987, USA 2: Department of Biology, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA 3: Department of Carcinogenesis, University of Texas M. D. Anderson Cancer Center, Smithville, Texas 78957, USA

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