Isolation and characterization of temperature-sensitive mutants of adenovirus type 7.
Fifty temperature-sensitive mutants, which replicate at 32 degrees C but not at 39.5 degrees C, were isolated after mutagenesis of the vaccine strain of adenovirus type 7 with hydroxylamine (mutation frequency of 9.0%) or nitrous acid (mutation frequency of 3.8%). Intratypic complementation analyses separated 46 of these mutants into seven groups. Intertypic complementation tests with temperature-sensitive mutants of adenovirus type 5 showed that the mutant in complementation group A failed to complement H5ts125 (a DNA-binding protein mutant), that mutants in group B and C did not complement adenovirus type 5 hexon mutants, and that none of the mutants was defective in fiber production. Further phenotypic characterization showed that at the nonpermissive temperature the mutant in group A failed to make immunologically reactive DNA-binding protein, mutants in groups B and C were defective in transport of trimeric hexons to the nucleus, mutants in groups D, E, and F assembled empty capsids, and mutants in group G assembled DNA-containing capsids as well as empty capsids. The mutants of the complementation groups were physically mapped by marker rescue, and the mutations were localized between the following map coordinates: groups B and C between 50.4 and 60.2 map units (m.u.), groups D and E between 29.6 and 36.7 m.u., and group G between 36.7 and 42.0 m.u. or 44.0 and 47.0 m.u. The mutant in group A proved to be a double mutant.
D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002213:Capsid / Q000032:analysis; D002460:Cell Line; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000032:analysis; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D005816:Genetic Complementation Test; D006801:Humans; D009154:Mutation; D010641:Phenotype; D013696:Temperature; D010948:Viral Plaque Assay
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D004262:DNA Restriction Enzymes