Ribosomal binding sites on poliovirus RNA

Eucaryotic ribosome binding sites on type 1 poliovirus RNA were obtained by isolation of T1 RNase-resistant RNA fragments from 80 S ribosomes that had been bound under conditions specific for initiation of translation and prevented from translocation with sparsomycin. The fragments were analyzed by two-dimensional polyacrylamide gel electrophoresis. Three ribosome-protected (RP) oligonucleotides and several non-ribosomal-protein-protected (NP) oligonucletides were analyzed. Secondary T1 digestion of the RP fragments revealed that at least three separate species existed. By comparison of these secondary digests to T1 digests of RNaseIII fragments mapped on the polio RNA, and by electron microscopic observation of ribosomes bound to polio RNA, preferred ribosome binding sites were localized to near 115 bases from the 5′ end, just to the 3′ side of midgenome, and 780 bases from the 3′ end.