Selection and preliminary characterization of temperature-sensitive mutants of type 5 adenovirus.
Eight temperature-sensitive (ts) mutants that replicate normally at 32 C but poorly, if at all, at 39.5 C have been isolated from mutagenized stocks of a wild-type strain of type 5 adenovirus. Three mutagens were employed: nitrous acid, hydroxylamine, and nitrosoguanidine. Ts mutants were isolated from mutagenized viral stocks with frequencies between 0.01 and 0.1%. All eight mutants had reversion frequencies of 10(-5) or less. Complementation experiments in doubly infected cultures at the nonpermissive temperature separated the mutants into three nonoverlapping complementation groups. Complementation yields ranged from a 2.3- to a 3,000-fold increase over the sums of the yields from the two singly infected controls. Genetic recombination was also demonstrated; approximate recombination frequencies ranged from 0.1 to 15%. Preliminary biochemical and immunological characterization of the mutants indicated that: (i) the single mutant in complementation group I did not replicate its deoxyribonucleic acid (DNA) or synthesize late proteins at the nonpermissive temperature but did inhibit host DNA synthesis to 25% of an uninfected control; (ii) the four group II mutants replicated viral DNA, shut off host DNA synthesis, synthesized penton base and fiber, but did not synthesize immunologically detectable hexon; the three mutants in complementation group III synthesized viral DNA, shut off host DNA synthesis, and made immunologically reactive capsid proteins (hexon, penton base, and fiber).
D000256:Adenoviridae / Q000254:growth & development* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000956:Antigens, Viral; D002247:Carbon Isotopes; D002277:Carcinoma; D002460:Cell Line / Q000382:microbiology; D004279:DNA, Viral / Q000096:biosynthesis; D005816:Genetic Complementation Test; D005827:Genetics, Microbial; D006801:Humans; D006898:Hydroxylamines; D005779:Immunodiffusion; D009062:Mouth Neoplasms; D009153:Mutagens; D009154:Mutation; D009573:Nitrites; D009604:Nitrosoguanidines; D011995:Recombination, Genetic; D012703:Serotyping; D013696:Temperature; D014316:Tritium; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000096:biosynthesis
D000956:Antigens, Viral; D002247:Carbon Isotopes; D004279:DNA, Viral; D006898:Hydroxylamines; D009153:Mutagens; D009573:Nitrites; D009604:Nitrosoguanidines; D014764:Viral Proteins; D014316:Tritium