Excess synthesis of viral mRNA 5-terminal oligonucleotides by reovirus transcriptase.

Abstract Short oligonucleotides corresponding to the 5'-terminal sequence of reovirus mRNAs were produced in vitro by virion-associated transcriptase activity. Both capped and uncapped oligonucleotides were synthesized in molar excess relative to mRNA. Yields of uncapped oligomers including ppG-C and ppG-C-U were severalfold greater than the homologous capped structures. In partial reaction mixtures that were nonsupportive for mRNA chain elongation, capped oligomer synthesis was increased. Similarly, oligonucleotide formation was differentially resistant in viral preparations that were inactivated with respect to mRNA synthesis by modification of the genome RNA by dimethyl sulfate alkylation or psoralen photoreaction. The results suggest that reovirus mRNA synthesis involves excessive initiation by reiterative transcription of promoter sites by the reovirus polymerase. Only a small fraction of the resulting oligonucleotides are capped and extended to form full length mRNAs during a subsequent elongation step which is apparently mediated by transcriptase molecules that escape the reiterative phase of transcription.