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PMID: 272636 Published · ppublish English Journal Article

Specific binding of messenger RNA and methionyl-tRNAfMet by the same initiation factor for eukaryotic protein synthesis.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 75 ·No. 1 ·1978-01-00 ·Pages 209-13

Kaempfer R, Hollender R, Abrams WR, Israeli R

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.

MeSH Terms
Base Sequence Binding Sites Chromatography, Affinity Guanosine Triphosphate/metabolism Methionine Nucleic Acid Conformation Peptide Initiation Factors/isolation & purification RNA, Messenger/metabolism RNA, Transfer/metabolism RNA, Viral/metabolism Ribosomes/metabolism
Chemicals
Peptide Initiation Factors RNA, Messenger RNA, Viral Guanosine Triphosphate RNA, Transfer Methionine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kaempfer R
Hollender R
Abrams W R
Israeli R
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27 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.75.1.209
Published
1978-01-00
Pages
209-13
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC411215
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.75.1.209 PMC Full Text View journal details
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