Identification of specific fragments containing the 5′ end of poliovirus RNA after ribonuclease III digestion

T.J.R. Harris, J.J. Dunn, E. Wimmer
1978 Nucleic Acids Research  
The small protein (VPg) covalently linked to the 5' end of poliovirus Type 1 (PV-1) RNA has been labeled in vitro with 125I using the Bolton and Hunter reagent. The RNA is not degraded under the conditions used and nearly all the label enters VPg and not the polynucleotide chain. When this 125I-labeled RNA is cleaved with RNase III at low monovalent salt concentrations, one major 125I-labeled fragment, approximately 100 nucleotides long, is produced. The corresponding fragment from similar
more » ... ts of 2P-labeled RNA has also been identified. The "2P-l abeled fragment changes electrophoretic mobility after protease treatment indicating that it contains VPg. Furthermore, the RNase Ti oligonucleotide known to be at the 5' terminus of poliovirus RNA is found in Ti digests of the purified fragment. These results confirm that the fragment is derived from the 5' end of the RNA. This fragment will be useful in studies concerning the initiation of protein synthesis during poliovirus infection.
doi:10.1093/nar/5.11.4039 pmid:214765 pmcid:PMC342732 fatcat:hkxieorqwnhfjf7knmz3fab6v4