Mapping Temperature-sensitive Mutants of Vesicular Stomatitis Virus by RNA Heteroduplex Formation

Authors:
  • GFG. J. Freeman
    G. J. Freeman
  • AHA. S. Huang
    A. S. Huang
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Abstract
Duplex RNA molecules made by hybridization of virion and mRNA of vesicular stomatitis virus (VSV) were digested with RNase and separated into 5 size classes, each containing the gene and the mRNA for 1 of the VSV proteins. Denaturation of the duplexes yielded full size mRNA lacking poly(A) tails. Utilizing duplex formation between the RNA from VSV temperature-sensitive (ts) mutants and their revertants and subsequent RNase digestion under varying salt conditions, specific cleavages within a certain duplex were seen for representative mutants from complementation groups III, IV and V. Specific cleavages were not seen for a group II mutant. Gene assignments cannot be made for group II; equivocal assignments are made for group III and clear assignments made for groups IV and V. The assignment for the group V mutants does not conform to expectations. Interactions may exist between the gene products of complementation groups II and V during VSV transcription and morphogenesis. The lack of transcriptional splicing for VSV mRNA is supported.

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