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Journal of Virology, November 2001, p. 11088-11095, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.11088-11095.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Comparison of Polymerase Subunits from Double-Stranded RNA Bacteriophages

Hongyan Yang, Eugene V. Makeyev, and Dennis H. Bamford*

Department of Biosciences and Institute of Biotechnology, FIN-00014 University of Helsinki, Finland

Received 16 April 2001/Accepted 16 August 2001

The family Cystoviridae comprises several bacteriophages with double-stranded RNA (dsRNA) genomes. We have previously purified the catalytic polymerase subunit (Pol) of one of the Cystoviridae members, bacteriophage phi 6, and shown that the protein can catalyze RNA synthesis in vitro. In this reaction, both bacteriophage-specific and heterologous RNAs can serve as templates, but those containing 3' termini from the phi 6 minus strands are favored. This provides a molecular basis for the observation that only plus strands, not minus strands, are transcribed from phi 6 dsRNA segments in vivo. To test whether such a regulatory mechanism is also found in other dsRNA viruses, we purified recombinant Pol subunits from the phi 6-related bacteriophages phi 8 and phi 13 and assayed their polymerase activities in vitro. The enzymes catalyze template-dependent RNA synthesis using both single-stranded-RNA (ssRNA) and dsRNA templates. However, they differ from each other as well as from phi 6 Pol in certain biochemical properties. Notably, each polymerase demonstrates a distinct preference for ssRNAs bearing short 3'-terminal sequences from the virus-specific minus strands. This suggests that, in addition to other factors, RNA transcription in Cystoviridae is controlled by the template specificity of the polymerase subunit.

* Corresponding author. Mailing address: Department of Biosciences and Institute of Biotechnology, P.O. Box 56, Viikinkaari 5, FIN-00014, University of Helsinki, Finland. Phone: 358 9 191 59100. Fax: 358 9 191 59098. E-mail: dennis.bamford{at}helsinki.fi.

Journal of Virology, November 2001, p. 11088-11095, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.11088-11095.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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