Ribosomal localization of the mRNA in the 30S initiation complex as revealed by UV crosslinking.
Translation initiation complexes consisting of 30S ribosomal subunits, 32P-labelled mRNA (002 mRNA), fMet-tRNA and the three initiation factors were subjected to UV-crosslinking to determine the protein and rRNA neighbors of the bound mRNA by immunochemical methods and by nucleic acid hybridization techniques. The mRNA was found to be crosslinked to a specific region of the 16S rRNA spanning from nucleotide 418 to 615 and to ribosomal proteins S1 and S21 (the main targets), S3, S10, S12 and S14; a low level of crosslinking was also detected with S2, S7, S13, S18 and S19.
Brandt
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Max-Planck-Institut für Molekulare Genetik, Berlin, Germany.
Gualerzi
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CO
D001483:Base Sequence; D003198:Computer Simulation; D003432:Cross-Linking Reagents; D004926:Escherichia coli / Q000378:metabolism; D015151:Immunoblotting; D046911:Macromolecular Substances; D008961:Models, Structural; D008969:Molecular Sequence Data; D010442:Peptide Chain Initiation, Translational; D010448:Peptide Initiation Factors / Q000032:analysis / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000032:analysis / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D012269:Ribosomal Proteins