Mapping polycyclic aromatic hydrocarbon and aromatic amine-induced DNA damage in cancer-related genes at the sequence level.
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.
Tang
Moon-shong
MS
Department of Environmental Medicine, New York University School of Medicine, 57 Old Forge Road, Tuxedo, NY 10987, USA. tang@env.med.nyu.edu
Pfeifer
Gerd P
GP
Denissenko
Mikhail F
MF
Feng
Zhaohui
Z
Hu
Wenwei
W
Pao
Annie
A
Zheng
Yi
Y
Zheng
Jessica B
JB
Li
Haiying
H
Chen
James X
JX
D024322:Amino Acids, Aromatic / Q000009:adverse effects*; D002471:Cell Transformation, Neoplastic; D002874:Chromosome Mapping; D004249:DNA Damage; D004260:DNA Repair; D004785:Environmental Pollutants / Q000009:adverse effects*; D004926:Escherichia coli / Q000235:genetics; D016158:Genes, p53 / Q000235:genetics*; D011905:Genes, ras / Q000235:genetics*; D016296:Mutagenesis; D011084:Polycyclic Aromatic Hydrocarbons / Q000009:adverse effects*; D016133:Polymerase Chain Reaction
D024322:Amino Acids, Aromatic; D004785:Environmental Pollutants; D011084:Polycyclic Aromatic Hydrocarbons