Nongenetic complementation in VSV: asymmetric contribution of the L proteins of each parent in the rescue of group I ts mutants

Abstract

Complementation group I temperature-sensitive mutants of vesicular stomatitis virus (VSV) are rescued at nonpermissive temperature by UV-irradiated virus. Rescue is a nongenetic process mediated by the structural L protein molecules of the parental irradiated virus. This is shown by action spectra analysis (V. Deutsch, B. Muel, and G. Brun (1977), Virology 77, 294-305), efficiency of rescue at doses high enough to inactivate every gene I, and rescue by irradiated defective-interfering short particles. Viral molecular synthesis at 39.6 degrees using the ts genomes as templates and stimulated by UV-irradiated virus is shown by gel electrophoresis of rescued virion proteins. Parental strain ts 053(I) is thermolabile in vitro while the thermostability of the rescued virions, genetically characterized as ts group I virus, is identical to that of helper wt or ts+ revertant. This suggests that some parental Lwt molecules are reincorporated in the rescued virions, together with newly synthesized Lts molecules. Efficiency variation of the rescue as a function of the multiplicity of infection of the UV-irradiated virions depends on the m.o.i. of the unirradiated ts I mutants. This result suggests that rescue depends on the concentration of the helper L molecules and also of that of the L initially bound to the ts template. That L of both parents contribute to rescue is supported by the observations that (1) rescue by UV-wt is strongly diminished after in vitro heating of thermolabile ts O53(I). (2) Intragenic rescue can be demonstrated: The helper activity of a given UV-ts I mutant is different according to the unirradiated ts I mutant used; the activity of helper L associated with different templates in intragenic heterologous combinations is higher or lower than its activity in the homologous combination (self-rescue control), instead of being equal as expected. (3) Efficiency of rescue by UV-wt also varies according to the ts I mutant. The initially bound Lts seems therefore to play a role important and different from that of the helper L. Contribution of both parent L to rescue seems thus to be qualitatively different.