Minekawa Y

Characterization of host-range and temperature-sensitive mutants of adenovirus type 5 with particular regard to transformation of a hamster embryo cell line (Nil)

Abstract Seventeen conditional lethal mutants (7 host-range and 10 temperature-sensitive) of adenovirus type 5 (Ad5) were classified by complementation test and characterized physiologically in viral-DNA synthesis, induction of cell DNA synthesis (in hamster kidney cells), capsid polypeptides production, and transformation of Nil cells (a hamster embryo cell line) under the restrictive conditions. Seven host-range ( hr ) mutants were divided into six groups by complementation test and into three classes by phenotypic characterization. Mutants assigned to class III (complementation groups D, E, F) were positive in viral-DNA synthesis and capsid polypeptides (hexon, penton base, fiber) production, and showed some degree of leakiness. Class II mutants (complementation groups B, C) were positive in viral-DNA synthesis with a small amount of capsid polypeptides production. Class I mutant (complementation group A) was an early mutant defective in viral-DNA synthesis but positive in induction of host-DNA synthesis. Transformation of Nil cells was observed with classes I and II mutants and not with class III mutants. Ten temperature-sensitive ( ts ) mutants were divided into seven complementation groups and into five classes by the available phenotypic criteria. Class V mutant (complementation group G) was positive in viral-DNA synthesis and capsid polypeptides production with extreme leakiness. Class IV mutants (complementation groups E, F) were positive in viral-DNA synthesis and capsid polypeptides production. Class III mutants (complementation groups C, D) were quite similar to class IV except for reduced hexon production. Class II mutants (complementation group B) were early mutants defective in viral-DNA synthesis but positive in induction of host-DNA synthesis. Class I mutants (complementation group A) were similar to class II but with a reduced degree of induction of host-DNA synthesis. Transformation of Nil cells was observed with classes II, III, and IV mutants and not with I and V mutants. In brief, the phenotypic characterization of hr and ts mutants in infection of hamster cells showed a good correlation between complementation grouping and the defective function. Transformation of Nil cells was observed with most groups of the mutants except for the apparently leaky late groups and one group of early mutants under the restrictive conditions.