Contrasting genome-wide distribution of 8-hydroxyguanine and acrolein-modified adenine during oxidative stress-induced renal carcinogenesis.
Oxidative stress is a persistent threat to the genome and is associated with major causes of human mortality, including cancer, atherosclerosis, and aging. Here we established a method to generate libraries of genomic DNA fragments containing oxidatively modified bases by using specific monoclonal antibodies to immunoprecipitate enzyme-digested genome DNA. We applied this technique to two different base modifications, 8-hydroxyguanine and 1,N6-propanoadenine (acrotein-Ade), in a ferric nitrilotriacetate-induced murine renal carcinogenesis model. Renal cortical genomic DNA derived from 10- to 12-week-old male C57BL/6 mice, of untreated control or 6 hours after intraperitoneal injection of 3 mg iron/kg ferric nitrilotriacetate, was enzyme digested, immunoprecipitated, cloned, and mapped to each chromosome. The results revealed that distribution of the two modified bases was not random but differed in terms of chromosomes, gene size, and expression, which could be partially explained by chromosomal territory. In the wild-type mice, low GC content areas were more likely to harbor the two modified bases. Knockout of OGG1, a repair enzyme for genomic 8-hydroxyguanine, increased the amounts of acrolein-Ade as determined by quantitative polymerase chain reaction analyses. This versatile technique would introduce a novel research area as a high-throughput screening method for critical genomic loci under oxidative stress.
Akatsuka
Shinya
S
Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
Aung
Than Tin
TT
Dutta
Khokon Kumar
KK
Jiang
Li
L
Lee
Wen-Hua
WH
Liu
Yu-Ting
YT
Onuki
Janice
J
Shirase
Tomoyuki
T
Yamasaki
Kyoko
K
Ochi
Hirotomo
H
Naito
Yuji
Y
Yoshikawa
Toshikazu
T
Kasai
Hiroshi
H
Tominaga
Yohei
Y
Sakumi
Kunihiko
K
Nakabeppu
Yusaku
Y
Kawai
Yoshichika
Y
Uchida
Koji
K
Yamasaki
Aiichi
A
Tsuruyama
Tatsuaki
T
Yamada
Yoshihiro
Y
Toyokuni
Shinya
S
D000171:Acrolein / Q000737:chemistry; D000225:Adenine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics; D045647:DNA Glycosylases / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D052138:Genes, Neoplasm / Q000235:genetics*; D016678:Genome / Q000235:genetics; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D047468:Immunoprecipitation; D007668:Kidney / Q000737:chemistry; D007680:Kidney Neoplasms / Q000235:genetics*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010084:Oxidation-Reduction; D018384:Oxidative Stress
C514955:1,N6-propanoadenine; D000911:Antibodies, Monoclonal; C024829:8-hydroxyguanine; D006147:Guanine; D000171:Acrolein; D004247:DNA; D045647:DNA Glycosylases; C485990:Ogg1 protein, mouse; D000225:Adenine