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Temperature-Sensitive Mutants of Adenovirus Type 12 Defective in Viral DNA Synthesis

Department of Biology, University of Akron, Akron, Ohio 44325

ABSTRACT

Ten temperature-sensitive (ts) mutants of adenovirus type 12 which produce plaques at 31 but not at 38.5 C have been isolated after mutagenesis with nitrosoguanidine or nitrous acid. The mutants have been classified into six separate complementation groups. DNA-DNA hybridizations have shown that at 38.5 C the ts 401 and 406 mutants of groups B and E, respectively, synthesized less than 10% of the normal level of viral DNA. The two mutants were also defective in the production of late proteins at the nonpermissive temperature, as shown by fluorescent-antibody tests and analysis by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Genetic recombination between the ts viruses 401 and 406 has been demonstrated; the recombination frequency for the wild-type virus production was 17.7%. Both mutants induced an increase in thymidine kinase activity at 38.5 C. Moreover, the two viral DNA-defective mutants shut off host DNA synthesis at the restrictive temperature. It is striking that at 38.5 C ts virus 401 transformed two to eight times more hamster cells than the wild-type virus, whereas ts virus 406 transformed at a frequency similar to the wild-type virus.