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J Virol. 1973 August; 12(2): 253-264
Copyright © 1973 American Society for Microbiology. All Rights Reserved.

RNA Synthesis by Vesicular Stomatitis Virus and a Small Plaque Mutant: Effects of Cycloheximide

¹ Department of Human Genetics, University of Michigan Medical School, Ann Arbor, Michigan 48104

ABSTRACT

The synthesis of viral RNA by wild-type vesicular stomatitis virus (L₁VSV) and a small, plaque-size mutant (S₂VSV) was studied in vitro and in chicken embryo (CE) and mouse L-cell cultures. Virus-specific RNA synthesized in CE or L cells infected with either L₁ or S₂VSV at low multiplicity was of the same size classes, 12 to 15S, 28S, and 38S. The major differences were in the proportion of RNA produced of each size class. L₁VSV always synthesized larger proportions of 38S RNA, and S₂VSV produced larger proportions of 12 to 15S RNA. Both S₂ and L₁VSV exhibited RNA transcriptase activity in vitro and in cell culture. The products of the in vitro reaction were the same, 12 to 15S for both. The products of the virion-associated transcriptase in CE or L-cell cultures in the presence of cycloheximide were also the same for both viruses but differed from the in vitro products in that 28S and 12 to 15S RNA were made. The effects of addition of cycloheximide at various times after infection demonstrated that new protein synthesis is required early (0-2 h) for both S₂ and L₁VSV to initiate and maintain the normal rate of viral RNA synthesis. However, the overall rate of RNA synthesis in L₁VSV infections became independent of protein synthesis after 2 h whereas the rate in S₂VSV infections did not. With either virus, synthesis of 38S RNA did not occur in the absence of protein synthesis. Moreover, continuous 38S RNA production required continuous protein synthesis. Production of 38S RNA ceased within 30 min after addition of cycloheximide to S₂^– or L₁VSV-infected CE or L cells that had already begun to synthesize the 38S form. The cycloheximide-induced cessation of 38S RNA synthesis was accompanied by a marked increase in production of 12 to 15S and 28S RNA in L₁VSV-infected cells, but no increase in synthesis of small RNA species occurred in S₂VSV-infected cells.

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