Open AccessCross-linking of Met-tRNAfto eIF-2β and to the ribosomal proteins S3a and S6 within the eukaryotic initiation complex, eIF-2-GMPPCP Met-tRNAf-small ribosomal subunit
Author(s) -
Peter Westermann,
Odd Nygård,
H Bielka
Publication year - 1981
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/9.10.2387
Subject(s) - biology , ribosomal rna , ribosomal protein , ribosome , eukaryotic large ribosomal subunit , transfer rna , protein subunit , eukaryotic small ribosomal subunit , biochemistry , 5.8s ribosomal rna , microbiology and biotechnology , rna , gene
In the quaternary initiation complex, eIF-2.GMPPCP.Met-tRNAf.40S ribosomal subunit, the Met-tRNAf can be cross-linked to the beta subunit of initiation factor eIF-2 as well as to ribosomal proteins S3a and S6 by treatment with the bifunctional reagent, diepoxybutane. Using 40S subunits, modified in advance with the heterobifunctional reagent, methyl-rho-azido-benzoylaminoacetimidate, Met-tRNAf is covalently bound to the same ribosomal proteins (S3a and S6) upon irradiation of the complex with ultraviolet light. Under both conditions proteins S3a and S6, together with a limited number of other ribosomal proteins, are covalently bound to 18S ribosomal RNA.