Specific binding of messenger RNA and methionyl-tRNAfMet by the same initiation factor for eukaryotic protein synthesis.
- 1 January 1978
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 75 (1) , 209-213
- https://doi.org/10.1073/pnas.75.1.209
Abstract
Affinity chromatography on columns containing globin mRNA, R17 phage mRNA, or double-stranded RNA linked to cellose is used to demonstrate unequivocally that the eukaryotic initiation factor (eIF-2) that forms a ternary complex with Met-tRNAf and GTP also binds tightly to these RNA species. Affinity chromatography of reticulocyte ribosomal wash yields over 100-fold purification of Met-tRNAf-binding factor. This factor is eluted as one of the most tightly bound proteins, and is active in protein synthesis even after passage over a column of double-stranded RNA-cellulose. eIF-2 binds mRNA and double-stranded RNA in distinctly different modes, protecting essentially all sequences in double stranded RNA, but very few in mRNA, against digestion with ribonuclease. Apparently, eIF-2 recognized the A conformation of double-stranded RNA, but not its sequence. By contrast, globin, Mengo virus, R17 and vesicular stomatitis virus mRNA are shown to possess a high-affinity binding site for eIF-2 that is absent in negative-strand RNA of vesicular stomatitis virus, an RNA that cannot serve as messenger. The results support the concept that eIF-2, the initiation factor that binds Met-tRNAf, recognizes an internal sequence in mRNA essential for protein synthesis.Keywords
This publication has 27 references indexed in Scilit:
- Phosphorylation of initiation factor eIF-2 and the control of reticulocyte protein synthesisCell, 1977
- Resistance of bacterial protein synthesis to double-stranded RNABiochemical and Biophysical Research Communications, 1974
- Sequence of Events in Initiation of Translation: A Role for Initiator Transfer RNA in the Recognition of Messenger RNAProceedings of the National Academy of Sciences, 1974
- Initiation of protein synthesis: Evidence for messenger RNA-independent binding of methionyl-transfer RNA to the 40 S ribosomal subunitJournal of Molecular Biology, 1973
- Inhibition of Cellular Protein Synthesis by Double-Stranded RNA: Inactivation of an Initiation FactorProceedings of the National Academy of Sciences, 1973
- Initiation of Eukaryotic Protein Synthesis: [Met-tRNAf.40S Ribosome] Initiation Complex Catalysed by Purified Initiation Factors in the Absence of mRNANature New Biology, 1973
- Protein Initiation in Eukaryotes: Formation and Function of a Ternary Complex Composed of a Partially Purified Ribosomal Factor, Methionyl Transfer RNA f , and Guanosine TriphosphateProceedings of the National Academy of Sciences, 1973
- Preparation and analysis of l-[35S]methionine labeled transfer ribonucleic acids from rabbit liverAnalytical Biochemistry, 1972
- Double-Stranded Poliovirus RNA Inhibits Initiation of Protein Synthesis by Reticulocyte LysatesProceedings of the National Academy of Sciences, 1971
- Ribonucleic acid synthesis of vesicular stomatitis virusVirology, 1970