L929 cells infected with temperature sensitive mutants of vesicular stomatitis virus: virus replication is necessary for induction of changes in membrane permeability.
Infection of L929 murine cells with vesicular stomatitis virus (VSV) results in inhibition of host protein synthesis and appearance of membrane alterations at a time when cells are still actively engaged in viral protein synthesis. VSV temperature-sensitive (ts) mutants have been used to explore the role(s) played by the virus-coded proteins in the genesis of these effects. Cells were infected with each of five ts mutants representing the known complementation groups of VSV Indiana serotype, and incubated at permissive (32 degrees C) and non-permissive temperatures (39 degrees C). Protein synthesis in the presence and absence of Hygromycin B (Hyg. B) was analyzed during virus infection via incorporation of 35S-methionine in acid-precipitable material and SDS-polyacrylamide gel electrophoresis. Data indicate that mutants belonging to groups I (L protein), II (NS protein) and IV (N protein) do not inhibit host protein synthesis and do not induce any membrane changes when grown at the non-permissive temperature. Mutants of group III (M protein) and V (G protein), instead, do inhibit cell protein synthesis and induce membrane changes also when grown at the non-permissive temperature; this suggests that these effects do not correlate with the biological activity of these proteins and their interaction with the cellular membrane. On the other hand, mutants exhibiting defective steps of nucleocapsid replication are apparently unable to induce these effects once more suggesting that virus replication per se is essential, as also indirectly shown by experiments employing cycloheximide to mimic shut-off.
di Francesco
P
P
Laboratory of Virology, Instituto Superiore di Sanità, Rome, Italy.
Sorrentino
V
V
Battistini
A
A
Curatola
A M
AM
Rossi
G B
GB
D000818:Animals; D002463:Cell Membrane Permeability; D002472:Cell Transformation, Viral; D003513:Cycloheximide / Q000494:pharmacology; D004261:DNA Replication; D006921:Hygromycin B / Q000494:pharmacology; D007700:Kinetics; D007739:L Cells / Q000382:microbiology; D051379:Mice; D009154:Mutation; D014176:Protein Biosynthesis / Q000187:drug effects; D013696:Temperature; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*; D014779:Virus Replication
D006921:Hygromycin B; D003513:Cycloheximide