TY - JOUR
T1 - Specific binding of messenger RNA and methionyl-tRNA(f)(Met) by the same initiation factor for eukaryotic protein synthesis
AU - Kaempfer, R.
AU - Hollender, R.
AU - Abrams, W. R.
AU - Israeli, R.
PY - 1978
Y1 - 1978
N2 - Affinity chromatography on columns containing globin mRNA, R17 phage mRNA, or double-stranded RNA linked to cellulose is used to demonstrate unequivocally that the eukaryotic initiation factor (eIF-2) that forms a ternary complex with Met-tRNA(f) and GTP also binds tightly to these RNA species. Affinity chromatography of reticulocyte ribosomal wash yields over 100-fold purification of Met-tRNA(f)-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 recognizes 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-tRNA(f), recognizes an internal sequence in mRNA essential for protein synthesis.
AB - Affinity chromatography on columns containing globin mRNA, R17 phage mRNA, or double-stranded RNA linked to cellulose is used to demonstrate unequivocally that the eukaryotic initiation factor (eIF-2) that forms a ternary complex with Met-tRNA(f) and GTP also binds tightly to these RNA species. Affinity chromatography of reticulocyte ribosomal wash yields over 100-fold purification of Met-tRNA(f)-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 recognizes 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-tRNA(f), recognizes an internal sequence in mRNA essential for protein synthesis.
UR - http://www.scopus.com/inward/record.url?scp=0017871023&partnerID=8YFLogxK
U2 - 10.1073/pnas.75.1.209
DO - 10.1073/pnas.75.1.209
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C2 - 272636
AN - SCOPUS:0017871023
SN - 0027-8424
VL - 75
SP - 209
EP - 213
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 1
ER -