The effect of 17beta-estradiol-DNA adducts on the replication of exon # 5 of the human suppressor gene p53.
Using a PCR technique, exon # 5 of the human tumor suppressor gene p53 was amplified and ligated into the pCRII vector and transformed into Escherichia coli INV alphaF' competent cells. The cloned exon # 5 was 184 bp long. Evidence is presented to show that after dimethyldioxirane epoxidation, 17beta-estradiol was able to form 17beta-estradiol-DNA adducts and to strongly inhibit the replication of the cloned exon # 5 producing smaller sizes of DNA fragments and introducing errors of incorporation at the 3'-end of the terminating DNAs. The errors occurred mainly at the clusters of the complementary 'G' and 'A' bases on the template strand DNA, presumably, the major sites where the 17beta-estradiol-DNA adducts were formed.
Department of Biomedical Sciences, University of Illinois College of Medicine at Rockford, 61107, USA. fuliyu@uic.edu
F L
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Yu
W Y
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Zheng
M Y
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Wang
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Bender
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Cheerva
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Miller
D001483:Base Sequence; D003001:Cloning, Molecular; D018736:DNA Adducts / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D004852:Epoxy Compounds / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology*; D005091:Exons; D016158:Genes, p53 / Q000187:drug effects*; D006801:Humans; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
D018736:DNA Adducts; D004852:Epoxy Compounds; D004958:Estradiol; C064455:dimethyldioxirane