Structural and functional analysis of separated strands of killer double-stranded RNA of yeast

The two strands of the M double-stranded RNA species from a killer strain of Saccharomyces cerevisiae have been separated, and the 3'-terminal sequences of these strands have been determined. The positive strand programs the synthesis of the putative killer toxin precursor (M-p32) in a rabbit reticulocyte in vitro translation system. Only the negative strand hybridizes to the positive polarity transcript (m) synthesized in vitro by the virion-associated transcriptase activity. Secondary structural analysis of the extreme 3'-terminus of the negative strand using S1 nuclease is consistent with the presence of a large stem and loop structure previously proposed on the basis of RNA sequence data. This structure, and a similar structure at the corresponding 5'-terminus of the positive strand, may have functional significance in vivo.