Cross-linking of mRNA to initiation factor eIF-3, 24 kDa cap binding protein and ribosomal proteins SI, S3/3a, S6 and S11 within the 48S pre-initiation complex

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Abstract
Native small ribosomal subunits from rabbit reticulocytes contain all initiation factors necessary for the formation of the mRNA-containing 48S pre-initiation complex. The complex formed in the presence of Met-tRNA f and 125 I-labelled globin mRNA was cross-linked with diepoxybutane, and the covalent mRNA-protein complexes were isolated under denaturating conditions. The proteins of the covalent complex were identified as the 110, 95 and 66/64 kDa subunits of eIF-3. In addition, the 24 kDa cap binding protein and the ribosomal proteins S1, S3/3a, S6 and S11 were found covalently linked to the mRNA. Ribosomal protein6 S3/3a and S6 were also involved in the ribosomal mRNA-binding domain of reticulocyte polysomes.