Screening procedure for complementation-dependent mutants of vesicular stomatitis virus

Abstract

To isolate new types of vesicular stomatitis virus (VSV) mutants, a four-stage screen was developed which identifies and characterizes mutants capable of complementing the defect in the VSV temperature-sensitive mutant tsGll. Two types of mutants of VSV, Indiana serotype, have been found by using the screen; they are new temperature-sensitive mutants which are, of necessity, not in complementation group I and mutants which do not produce plaques under conditions of single infection at 31 C (the normal permissive temperature) and are, therefore, called complementation-dependent mutants. The newly isolated, temperature-sensitive mutants fall into three complementation groups, two of which are congruent with known complementation groups; the newly identified group extends to six the number of complementation groups of VSV Indiana. The nature of the complementation-dependent mutants has not been established, but one was shown to not contain a significant deletion in its nucleic acid.

Selective isolation of mutants of vesicular stomatitis virus defective in production of the viral glycoprotein

“…(i) 5'-Fluorouracil and 5-azacytidine (16,20). Cultures of Vero cells were infected with a multiplicity of 0.5 of VSV and were grown for 8 h at 32°C in the presence of 0.30 mg of 5-fluorouracil (FU) or 0.004 mg of 5-azacytidine per ml.…”

“…VSV muta-tions defective in glycoprotein biosynthesis are rare; indeed, Pringle characterized over 200 fluorouracil-induced temperature-sensitive (ts) mutations in the Indiana strain of VSV and found none in the complementation group (V) for the G gene (16)(17)(18). At least five ts mutations in group V are known (4,20). In the case of the two mutants (ts045, tsM501) that have been studied most intensively, at the nonpermissive temperature G protein is synthesized and inserted into the rough endoplasmic reticulum (rer) (10).…”

Analysis of the defects of temperature-sensitive mutants of vesicular stomatitis virus: intracellular degradation of specific viral proteins

“…Temperature-sensitive mutants of vesicular stomatitis virus (VSV) have been isolated in several laboratories and classified into six complementation groups (2,3,6,18,19,21). Group I contains more than one-half of the mutants isolated, and some of these are defective for transcription of mRNA species in vivo at the nonpermissive temperature (15,16,20,23).…”

Virion trascriptase activity differences in host range mutants of vesicular stomatitis virus

“…Temperature-sensitive (ts) mutants of VSV Indiana fall into six complementation groups (5,8), and one of these groups is now considered to represent the cistron coding for the transcriptase L protein (3,6). Comparison of representative mutants from five groups showed certain mutants in group I to be irreversibly inactivated by incubation at 39 C. The thermolability of these mutants was considered to be a direct consequence of heat inactivation of the transcriptase (10).…”