Abstract
Sites and types of mutations in relation to the amount of cyclobutane pyrimidine dimers (CPDs) and (6-4)-photoproducts in UV-irradiated normal Ha-ras sequences were investigated. Mouse BALB/C 3T3 cells were transfected with UV-irradiated pYN-mHras plasmids containing mouse normal Ha-ras sequences, and transformed foci developed. Direct DNA sequencing of the Ha-ras retrieved from the foci revealed that most mutations (23/24, 96%) took place at dipyrimidine sequences, and the C-->T transition at the 3'-cytosine in 5'-TC or 5'-CC sequences was predominant (17/24, 71%) in codons 12, 13 and 60. In codon 61, where 5'-TC or 5'-CC is absent, two mutations were found at the 5'-TT sequence. More (6-4)photoproducts were produced than CPDs in codons 12, 13 and 60, and more CPDs were produced than (6-4)photoproducts in codon 61. These results suggest that (6-4)photoproducts are the major lesion leading to the mutations in the mouse Ha-ras sequence and subsequent transformation of BALB/C 3T3 cells.
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