TY - JOUR
T1 - Butadiene-induced intrastrand DNA cross-links
T2 - A possible role in deletion mutagenesis
AU - Carmical, J. Russ
AU - Kowalczyk, Agnieszka
AU - Zou, Yue
AU - Van Houten, Bennett
AU - Nechev, Lubomir V.
AU - Harris, Constance M.
AU - Harris, Thomas M.
AU - Lloyd, R. Stephen
PY - 2000/6/30
Y1 - 2000/6/30
N2 - To initiate studies designed to identify the mutagenic spectrum associated with butadiene diepoxide-induced N2-N2 guanine intrastrand cross-links, site specifically adducted oligodeoxynucleotides were synthesized in which the adducted bases were centrally located within the context of the human ras 12 codon. The two stereospecifically modified DNAs and the corresponding unmodified DNA were ligated into a single-stranded M13mp7L2 vector and transfected into Escherichia coli. Both stereoisomeric forms (R,R and S,S) of the DNA cross-links resulted in very severely decreased plaque-forming ability, along with an increased mutagenic frequency for both single base substitutions and deletions compared with unadducted DNAs, with the S,S stereoisomer being the most mutagenic. Consistent with decreased plaque formation, in vitro replication of DNA templates containing the cross-links by the three major E. coli polymerases revealed replication blockage by both stereoisomeric forms of the cross-links. The same DNAs that were used for replication studies were also assembled into duplex DNAs and tested as substrates for the initiation of nucleotide excision repair by the E. coli UvrABC complex, UvrABC incised linear substrates containing these intrastrand cross-links with low efficiency, suggesting that these lesions may be inefficiently repaired by the nucleotide excision repair system.
AB - To initiate studies designed to identify the mutagenic spectrum associated with butadiene diepoxide-induced N2-N2 guanine intrastrand cross-links, site specifically adducted oligodeoxynucleotides were synthesized in which the adducted bases were centrally located within the context of the human ras 12 codon. The two stereospecifically modified DNAs and the corresponding unmodified DNA were ligated into a single-stranded M13mp7L2 vector and transfected into Escherichia coli. Both stereoisomeric forms (R,R and S,S) of the DNA cross-links resulted in very severely decreased plaque-forming ability, along with an increased mutagenic frequency for both single base substitutions and deletions compared with unadducted DNAs, with the S,S stereoisomer being the most mutagenic. Consistent with decreased plaque formation, in vitro replication of DNA templates containing the cross-links by the three major E. coli polymerases revealed replication blockage by both stereoisomeric forms of the cross-links. The same DNAs that were used for replication studies were also assembled into duplex DNAs and tested as substrates for the initiation of nucleotide excision repair by the E. coli UvrABC complex, UvrABC incised linear substrates containing these intrastrand cross-links with low efficiency, suggesting that these lesions may be inefficiently repaired by the nucleotide excision repair system.
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U2 - 10.1074/jbc.M002037200
DO - 10.1074/jbc.M002037200
M3 - Article
C2 - 10766753
AN - SCOPUS:0034733643
SN - 0021-9258
VL - 275
SP - 19482
EP - 19489
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 26
ER -