Nucleic Acids Research, 1984, Vol. 12, No. 23 8887-8897
© 1984
MOLECULAR BIOLOGY |
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
Central Institute of Molecular Biology, Academy of Sciences of GDR 1115 Berlin-Buch, GDR 1The Wenner-Gren Institute, University of Stockholm S-113 45 Stockholm, Sweden
*To whom correspondence should be addressed.
Received September 17, 1984. Revised November 6, 1984. Accepted November 6, 1984.
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-tRNAf and 125I-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.
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