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Nucleic Acids Research, 1980, Vol. 8, No. 11 2349-2364
© 1980

Articles

Virion DNA-independent RNA polymerase from Saccharomyces cerevisiae

J.Douglas Welsh*, Michael J. Leibowitz* and Reed B. Wicknert{dagger}

*Department of Microbiology, CMDNJ-Rutgers Medical School Piscataway, NJ 08854, USA {dagger}Laboratory of Biochemical Pharmacology, National Institute of Arthritis, Metabolic and Digestive Diseases NIH, Bethesda, MD 20014, USA

Received May 7, 1980. The "killer" plasmid and a larger double-stranded RNA plasmid of yeast exist in intracellular virion particles. Purification of these particles from a diploid killer strain of yeast (grown into stationary growth on ethanol) resulted in co-purification of a DNA-independent RNA polymerase activity. This activity incorporates and requires all four ribonucleoside triphosphates and will not act on deoxyribonucleoside triphosphates. The reaction requires magnesium, is inhibited by sulfhydryl-oxidizing reagents and high concentrations of monovalent cation, but is insensitive to DNase, {alpha}-amanitin, and actinomycin D. Pyrophosphate inhibits the reaction as does ethidium bromide. Exogenous nucleic acids have no effect on the reaction. The product is mostly single-stranded RNA, some of which is released from the enzymatically active virions.


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