Transcription of vesicular stomatitis virus (VSV) in Vero cells was confined to the synthesis of parentally-derived mRNA (primary transcription) by the use of cycloheximide and/or a ts mutant, G41(IV), at a non-permissive temperature (40 °C). More transcripts accumulated in the presence of cycloheximide than in its absence. This so-called ‘cycloheximide effect’ results from higher rates of virus transcription sustained for longer periods of time. The rate of VSV transcription initially increases linearly for 1 to 2 h after infection. Interferon reduces this rate (≃ fourfold with 50 units/ml interferon) irrespective of the presence or absence of cycloheximide. The VSV mRNA transcripts synthesized in mock- or interferon-treated cells were equal in size and had an equivalent half-life of 17 h at 40 °C. It seems likely that once transcription is initiated in interferon-treated cells, it is completed successfully.
Since interferon reduces the rate of early VSV primary transcript synthesis to below that achieved in the presence of cycloheximide, we conclude that interferon has an effect on transcription beyond that attributable solely to protein synthesis inhibition. We postulate that interferon decreases the probability of initiating virus transcription. Virus mRNA escaping this facet of interferon action may then encounter other facets such as post-transcriptional modification and/or inhibition of translation. However, the mandatory sequence of primary transcription → primary translation for negative-strand viruses like VSV dictates that the overall inhibitory effect of interferon on translation would derive in part from this prior inhibition of transcription. Thus, to apply the term ‘primary effect’ to one particular facet of interferon action may not always be meaningful.
BaltimoreD.,
HuangA. S.,
StampferM.1970; Ribonucleic acid synthesis of vesicular stomatitis virus. II. An RNA polymerase in the virion. Proceedings of the National Academy of Sciences of the United States of America 66:572–576
BaxtB.,
SonnabendJ. A.,
BablanianR.1977; Effects of interferon on vesicular stomatitis virus transcription and translation. Journal of General Virology 35:325–334
BrownG. E.,
LebleuB.,
KawakitaM.,
ShailaS.,
SenG. C.,
LengyelP.1976; Increased endonuclease activity in an extract from mouse Ehrlich ascites tumour cells which had been treated with a partially purified interferon preparation: dependence on double-stranded RNA. Biochemical and Biophysical Research Communications 69:114–122
CombardA.,
MartinetC.,
Printz-AneC.,
FriedmanA.,
PrintzP.1974; Transcription and replication of VSV: effects of temperature-sensitive mutations in complementation group IV. Journal of Virology 13:922–930
EstebanM.,
MetzD. H.1973; Inhibition of early vaccinia virus protein synthesis in interferon-treated chicken embryo fibroblasts. Journal of General Virology 20:111–115
FlamandA.,
BishopD. H. L.1973; Primary in vivo transcription of VSV and temperature-sensitive mutants of five VSV complementation groups. Journal of Virology 12:1238–1252
GaunttC. J.1972; Effect of interferon synthesis on ssRNA in reovirus type 3-infected L-cell cultures. Biochemical and Biophysical Research Communications 47:1228–1236
GentyN.1975; Analysis of uridine incorporation in chicken embryo cells infected by vesicular stomatitis virus and its temperature-sensitive mutants: uridine transport. Journal of Virology 15:8–15
GravellM.,
CromeansT. L.1972; Inhibition of early viral RNA synthesis in interferon-treated cells infected with frog polyhedral cytoplasmic deoxyribovirus. Virology 50:916–919
HuangA. S.,
MandersE. R.1972; RNA synthesis of vesicular stomatitis virus. IV. Transcription by standard virus in the presence of defective interfering particles. Journal of Virology 9:909–916
HuntD. M.,
WagnerR. R.1974; Location of the transcription defect in Group I temperature-sensitive mutants of vesicular stomatitis virus. Journal of Virology 13:28–35
JoklikW. K.,
MeriganT. C.1966; Concerning the mechanism of action of interferon. Proceedings of the National Academy of Sciences of the United States of America 56:558–565
KatesJ. R.,
McauslanB. R.1967; Messenger RNA synthesis by a ‘coated’ viral genome. Proceedings of the National Academy of Sciences of the United States of America 57:314–320
KileyM. P.,
PayneF. E.1974; Replication of measles virus: continued synthesis of nucleocapsid RNA and increased synthesis of mRNA in the presence of cycloheximide. Journal of Virology 14:758–764
MarcusP. I.,
SekellickM. J.1974; Cell-killing by viruses. I. Comparison of cell-killing, plaque-forming and defective-interfering particles of vesicular stomatitis virus. Virology 57:321–338
MarcusP. I.,
SekellickM. J.1975; Cell-killing by viruses. II. Cell-killing by vesicular stomatitis virus: a requirement for virion-derived transcription. Virology 63:176–190
MarcusP. I.,
SekellickM. J.1976; Cell-killing by viruses. III. The interferon system and inhibition of cell-killing by vesicular stomatitis virus. Virology 69:378–393
MarcusP. I.,
SekellickM. J.,
JohnsonL. D.,
LazzariniR. A.1977; Cell-killing by viruses. V. Transcribing defective interfering particles of vesicular stomatitis virus function as cell-killing particles. Virology 82:242–246
MarcusP. L.,
TerryT. M.,
LevineS.1975; Interferon action II. Membrane-bound alkaline ribonuclease activity in chick embryo cells manifesting interferon-mediated interference. Proceedings of the National Academy of Sciences of the United States of America 72:182–186
MarvaldiJ. L.,
Lucas-LenardJ.,
SekellickM. J.,
MarcusP. l.1977; Cell-killing by viruses IV. Cellkilling and protein synthesis inhibition by vesicular stomatitis virus require the same gene functions. Virology 79:267–280
MayG.,
NaglerS.,
GraziadeiW.1975; Aberrant species of single-stranded RNA synthesized by reovirus parental subviral particles isolated from interferon treated L-cells. American Society of Microbiology Abstracts p 239
MeeganJ. M.,
FullerF. J.,
MarcusP. I.1976; Poly(rI) Poly(rC) induces a double-stranded ribonuclease (dsRNase) in avian cells. American Society of Microbiology Abstracts p 238
MetzD. H.,
LevinM. J.,
OxmanM. N.1976; Mechanism of interferon action: further evidence for transcription as the primary site of action in Simian Virus 40 infection. Journal of General Virology 32:227–240
OxmanM. N.,
LevinM. J.1971; Interferon and transcription of early virus-specific RNA in cells infected with simian virus 40. Proceedings of the National Academy of Sciences of the United States of America 68:299–302
PerlmanS.M.,
HuangA. S.1973; RNA synthesis of vesicular stomatitis virus. V. Interactions between transcription and replication. Journal of Virology 12:1395–1400
PringleC. R.1970; Genetic characteristics of conditional lethal mutants of VSV induced by 5-fluorouracil, 5-azacytidine, and ethyl methane sulfonate. Journal of Virology 5:559–567
RepikP.,
FlamandA.,
BishopD. H. L.1974; Effect of interferon upon the primary and secondary transcription of VSV and influenza viruses. Journal of Virology 14:1169–1178
SzilagyiJ. F.,
PringleC. R.1972; Effect of temperature-sensitive mutations on the virion-associated RNA transcriptase of vesicular stomatitis virus. Journal of Molecular Biology 71:281–291
WertzG. W.1975; Method of examining viral RNA metabolism in cells in culture: metabolism of vesicular stomatitis virus RNA. Journal of Virology 16:1340–1344
WertzG. W.,
LevineM.1973; RNA synthesis by vesicular stomatitis virus and a small plaque mutant: effects of cycloheximide. Journal of Virology 12:253–264