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Volume 66, Issue 5

The Role of Polypeptides L and NS in the Transcription Process of Vesicular Stomatitis Virus New Jersey Using the Temperature-sensitive Mutant E1 Free

Abstract

SUMMARY

The roles of the L and NS polypeptides in transcription by vesicular stomatitis virus New Jersey were studied using a mutant, E1, which contains a temperature-sensitive transcriptase and an altered NS polypeptide, both phenotypic changes being the consequence of the mutation. Mutant E1, its revertant (E1/R1) and the wild-type virus were dissociated into sub-viral fractions and, after reconstitution of these fractions in all combinations, the transcriptase was assayed at the permissive (31 °C) and restrictive (39 °C) temperatures. Reconstitution of the pellet fractions (containing polypeptide N complexed with the virion RNA) and the supernatant fractions (containing polypeptides L and NS) restored transcriptase activity at 31 °C in all combinations, but at 39 °C transcription was observed only in the presence of the supernatant fractions of wild-type and revertant viruses but not in the presence of the supernatant fractions of E1. When the pellet fractions and the L fractions were reconstituted, the transcriptase activity was restored in all combinations both at 31 °C and 39 °C. However, transcription at 39 °C by reconstituted pellet and L fractions was strongly inhibited when the NS fraction of E1 was also added, while addition of the NS fractions of wild-type and revertant viruses had no effect. Since only traces of polypeptide NS were present in the L fractions and none in the pellet fractions, the results strongly suggest that polypeptide L is the transcriptase itself while polypeptide NS exerts some control over transcription.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): dissociation-reconstitution , mutant tsE1 , transcription in vitro and VSV
© Journal of General Virology 1985
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1985-05-01
2024-05-03
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The Role of Polypeptides L and NS in the Transcription Process of Vesicular Stomatitis Virus New Jersey Using the Temperature-sensitive Mutant tsE1
J. Gen. Virol. 66, 1011 (1985); https://doi.org/10.1099/0022-1317-66-5-1011
/content/journal/jgv/10.1099/0022-1317-66-5-1011
/content/journal/jgv/10.1099/0022-1317-66-5-1011
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