RNA Polymerase Activity Associated with Bacteriophage φ6

Authors

James L. Van Etten, University of Nebraska-LincolnFollow
A. K. Vidaver, University of Nebraska-LincolnFollow
R. K. Koski, University of Nebraska-Lincoln
J. S. Semancik, University of Nebraska-Lincoln

Date of this Version

9-1-1973

Comments

Published in JOURNAL OF VIROLOGY, Sept. 1973, p. 464-471 Vol. 12, No. 3. Copyright © 1973 American Society for Microbiology. Used by permission.

Abstract

The Pseudomonas phaseolicola bacteriophage φ6 incorporated labeled UTP into an acid-insoluble precipitate. Incorporation was dependent on the presence of manganese acetate, ATP, GTP, CTP, and a short heat treatment of the phage; the reaction was stimulated by NH,Cl. The substitution of ¹⁴C-ATP, -CTP or -GTP for UTP, together with the appropriate unlabeled ribonucleoside triphosphates, disclosed that CMP was incorporated to the greatest extent followed by GMP, UMP, and AMP. Radioactive RNAs formed by the reaction were resistant to RNases A and T₁ in high salt but susceptible to these nucleases in low salt. The labeled RNA co-sedimented and co-electrophoresed with φ6 double-stranded (ds) RNA. However, the distribution of the radioactivity into the three ds-RNA components varied depending on the ¹⁴4C-ribonucleoside triphosphate used in the reaction. The incorporation of UMP was primarily into the two smaller ds-RNA segments, GMP primarily into the large ds-RNA segment, and CMP and AMP were about equally distributed into all three ds-RNA segments.