Involvement of eukaryotic initiation factor 4A in the cap recognition process

• Published in: The Journal of biological chemistry Vol. 258; no. 18; pp. 11398 - 11403
• Main Authors: Edery, I, Hümbelin, M, Darveau, A, Lee, K A, Milburn, S, Hershey, J W, Trachsel, H, Sonenberg, N
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Elsevier Inc 25-09-1983; American Society for Biochemistry and Molecular Biology
• Subjects: Adenosine Triphosphate - metabolism; Animals; Biological and medical sciences; Carrier Proteins - isolation & purification; Chromatography, Affinity; Eukaryotic Initiation Factor-4A; Fundamental and applied biological sciences. Psychology; Molecular and cellular biology; Molecular genetics; Molecular Weight; Peptide Fragments - analysis; Peptide Initiation Factors - metabolism; Rabbits; RNA Cap-Binding Proteins; RNA Caps - metabolism; Translation. Translation factors. Protein processing; Trypsin - metabolism; Binding protein; Initiation factor eIF4A; Messenger RNA; Cap structure; Molecular interaction; Monoclonal antibody; Comparative study; Immunological method
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)44431-0

Antibodies against eukaryotic initiation factor 4A (eIF-4A) were used to study the involvement of this factor in recognizing the 5‘ cap structure of eukaryotic mRNA. We demonstrate that an approximately 50-kilodalton polypeptide present in rabbit reticulocyte ribosomal high salt wash which can be specifically cross-linked to the 5‘ oxidized cap structure of reovirus mRNA (Sonenberg, N. (1981) Nucleic Acids Res. 9, 1643) reacts with an anti-eIF-4A monoclonal antibody. We also show that antibodies against eIF-4A react with a 50-kilodalton polypeptide present in a cap-binding protein complex obtained by elution from a m7GTP-agarose affinity column. Comparative peptide analysis of eIF-4A and the 50-kilodalton component of the cap-binding protein complex indicates a very strong similarity between the two polypeptides.