Inhibition of translation in lysates of mouse L cells infected with vesicular stomatitis virus: presence of a defective ribosome-associated factor.
Lysates of L cells infected for 4 h with vesicular stomatitis virus were inhibited in their in vitro translational activity to about the same extent as protein synthesis was inhibited in vivo in infected L cells. Inhibition of translation occurred at the level of the ribosome as determined by reciprocal cross-reconstitution studies with polyribosomes and postribosomal supernatant fractions isolated from virus-infected and mock-infected cells. Inhibition of protein synthesis in reconstituted lysates of virus-infected cells was found to be at the level of initiation of translation as evidenced by reduction in incorporation into acid-precipitable proteins of formylatable [35S]methionine. Ribosomes from virus-infected and mock-infected cells were exposed to 0.5 M KCl and fractionated by centrifugation into salt-washed polyribosomes and supernatant fractions containing ribosome-associated proteins; reciprocal reconstitution of translational activity by a mixing of salt-washed polyribosomes and ribosome-associated proteins revealed that the defect in initiation of translation was in the ribosome-associated proteins released by salt wash from the infected-cell ribosomes. Differential ammonium sulfate precipitation of the supernatant ribosome-associated proteins from virus-infected and mock-infected cells indicated by reciprocal reconstitution studies that the defective ribosomal initiation factor(s) was (were) present primarily in the 0-40% ammonium sulfate fraction that is considered to contain primarily eIF-3 and eIF-4B. These results are similar to those found in earlier studies of defective initiation factors responsible for impaired protein synthesis in cells infected with plus-strand viruses quite different from the rhabdovirus studied in these experiments.
D000818:Animals; D005347:Fibroblasts / Q000382:microbiology; D051379:Mice; D010448:Peptide Initiation Factors / Q000032:analysis*; D014176:Protein Biosynthesis / Q000187:drug effects*; D012270:Ribosomes / Q000032:analysis*; D014721:Vesicular stomatitis Indiana virus
D010448:Peptide Initiation Factors