W16B is a temperature-sensitive mutant of vesicular stomatitis virus. Others have shown that it is temperature-sensitive for replication and for transcription and that these phenotypes are probably due to mutation of the N (nucleocapsid) gene. Five independent revertants were isolated from W16B based on their ability to grow at 39 °C. The thermosensitivity of transcription by these revertants was similar to that of the wild-type virus [(HR)] from which W16B was derived. Fractionation-reconstitution studies of two revertants indicated that the reversion was in the N or P (phosphoprotein) gene. The N and P genes of (HR), W16B, and these two revertants were sequenced. There were no differences between the P genes. Comparison of the predicted N protein sequences of (HR), W16B and the two revertants indicated that the growth and transcription phenotypes of W16B were due to a change of amino acid residue 238 from threonine to isoleucine. The amino acid at position 238 in the other three revertants also showed an exact reversion to threonine. Amino acid residue 238 lies in a domain of the N protein which is highly conserved among vesiculoviruses.


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