Mechanism of mutation on DNA templates containing synthetic abasic sites: study with a double strand vector.
Mutagenesis at abasic sites was investigated in E.coli and simian kidney (COS) cells using a duplex shuttle vector containing synthetic analogs of deoxyribose on the phosphodiester backbone. Lesions were positioned on opposite strands of the vector. When the tetrahydrofuranyl analog was used as the abasic site, AT or TA pairs (65-80%) were introduced at the site of the bistrand lesion. Mutagenesis occurred in the absence of SOS induction. Single base deletions (> 80%) dominated the mutational spectra for propanyl and ethanyl analogs of abasic sites lacking a ring structure. For all abasic site analogs, a small proportion of G/C and C/G pairs (6-10%) were observed. dAMP was incorporated predominantly opposite tetrahydrofuranyl sites positioned in the single strand region of a gapped duplex vector. We conclude from these studies that abasic sites positioned in a bistrand configuration are highly mutagenic in E.coli and COS cells. Repair DNA synthesis may be involved in this process.
Takeshita
M
M
State University of New York at Stony Brook, Department of Pharmacological Sciences 11794-8651.
Eisenberg
W
W
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000378:metabolism; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D003855:Deoxyribose / Q000031:analogs & derivatives; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000502:physiology*; D011089:Polydeoxyribonucleotides / Q000138:chemical synthesis; D013698:Templates, Genetic; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D003838:Deoxyadenine Nucleotides; D011089:Polydeoxyribonucleotides; D003855:Deoxyribose; C012873:2'-deoxy-5'-adenosine monophosphate; D004247:DNA