title: How are tRNAs and mRNA arranged in the ribosome? An attempt to correlate the stereochemistry of the tRNA-mRNA interaction with constraints imposed by the ribosomal topography.
abstract: Two tRNA molecules at the ribosomal A- and P-sites, with a relatively small angle between the planes of the L-shaped molecules, can be arranged in two mutually exclusive orientations. In one (the 'R'-configuration), the T-loop of the A-site tRNA faces the D-loop of the P-site tRNA, whereas in the other (the 'S'-configuration) the D-loop of the A-site tRNA faces the T-loop of the P-site tRNA. A number of stereochemical arguments, based on the crystal structure of 'free' tRNA, favour the R-configuration. In the ribosome, the CCA-ends of the tRNA molecules are 'fixed' at the base of the central protuberance (the peptidyl transferase centre) of the 50S subunit, and the anticodon loops lie in the neck region (the decoding site) of the 30S subunit. The translocation step is essentially a rotational movement of the tRNA from the A- to the P-site, and there is convincing evidence that the A-site must be located nearest to the L7/L12 protuberance of the 50S subunit. The mRNA in the two codon-anticodon duplexes lies on the 'inside' of the 'elbows' of the tRNA molecules (in both the S-type and R-type configurations), and runs up between the two molecules from the A- to the P-site in the 3' to 5'-direction. These considerations have the consequence that in the S-configuration the mRNA in the codon-anticodon duplexes is directed towards the 50S subunit, whereas in the R-configuration it is directed towards the 30S subunit. The results of site-directed cross-linking experiments, in particular cross-links to mRNA at positions within or very close to the codons interacting with A- or P-site tRNA, favour the latter situation. This conclusion is in direct contradiction to other current models for the arrangement of mRNA and tRNA on the ribosome.
authors.affiliation: Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region.
authors.forename: V
authors.initials: V
authors.lastname: Lim
authors.forename: C
authors.initials: C
authors.lastname: Venclovas
authors.forename: A
authors.initials: A
authors.lastname: Spirin
authors.forename: R
authors.initials: R
authors.lastname: Brimacombe
authors.forename: P
authors.initials: P
authors.lastname: Mitchell
authors.forename: F
authors.initials: F
authors.lastname: Müller
mesh_terms: D003432:Cross-Linking Reagents; D004926:Escherichia coli; D046911:Macromolecular Substances; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D010444:Peptide Elongation Factor Tu / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism / Q000648:ultrastructure*; D012343:RNA, Transfer / Q000378:metabolism / Q000648:ultrastructure*; D012270:Ribosomes / Q000378:metabolism / Q000648:ultrastructure*; D013237:Stereoisomerism
chemical_list: D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D012333:RNA, Messenger; D012343:RNA, Transfer; D010444:Peptide Elongation Factor Tu