5′-Terminal phosphorylation and secondary structure of double-stranded RNA from a fungal virus

Abstract

Different double-stranded RNA species from Penicillium stoloniferum virus have been phosphorylated at the 5′ termini with the aid of polynucleotide kinase. A very low phosphate uptake has been observed which, especially in the case of a relatively small molecular component, was increased several times by pretreatment with RNAase t 1. Adenosine and uridine have been detected at the 5′-termini of this RNA component. Digestion with RNAase T 1, an enzyme which does not cut across the two strands of a double-stranded RNA molecule, produced a new uridine terminus and increased the efficiency of phosphorylation. It is concluded that this double-stranded RNA molecule contains single-stranded stretches at or near the 5′-termini. The possibility of a circular structure being formed by the annealing of single-stranded tails is discussed.