Reovirus genome RNA segments: Resistance to S 1 nuclease

Abstract

Reovirion genome RNA was purified by sedimentation in a sucrose-formamide gradient after treatment with 50% formamide under nondenaturing conditions for genome RNA. Both the 5′- and 3′-termini of purified double-stranded (ds) RNA were completely resistant to digestion by the single-stranded specific nuclease from Aspergillus oryzae, S 1. In contrast, the unpaired 3′-terminal dinucleotide, … CpC- 3H, in tRNA was completely hydrolyzed by the enzyme. Reovirus ds RNA was sensitive to T 1 and pancreatic RNases to a small extent, even in the presence of 0.3 M NaCl. Ds RNA did not act as a primer/template for avian myeloblastosis virus (AMV) reverse transcriptase. The results indicate that reovirus ds genome RNA does not contain S 1-sensitive single-stranded regions.