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Translation III Elongation - Big Picture Ribosome selects the aa-tRNA in the A-site according to the sequence of the codons (decoding) Ribosome catalyzes peptide-bond formation between the C-terminus of the peptide attached to the tRNA in the P-site and the new aa bound to the tRNA in the A-site - simultaneous movement of large particle relative to mRNA and tRNAs by exactly one codon Movement of the tRNAs and mRNA relative to the small particle by one site Errors in translocation can lead to fameshifting and wrong protein products Recap: What are A, P and E sites on ribosome Steps of elongation Role of elongation factors in prokaryotes EF1A (EF-Tu) G protein(GTP/GDP-binding protein) Active when GTP is bound Kd = 10-6 M inactive with GDP is bound Kd = 10-8 M, slow offrate Offrate is accelerated by EF1B EF1B (EF-Ts) Guanosine nucleotide exchange factor (GEF) for EF1A(EF-Tu) EF2 (EF-G) G protein There is no known GEF for EF-G More of Elongation Factors EF1A (EF-Tu) 44 kDa protein, three domains Similar to eukaryotic eEF1A Function is to bind aa-tRNA protect aa-ester bond from hydrolysis Ternary complex of EF1A (EF1A/GTP/aa-tRNA) assists in binding aa-tRNA to the ribosome Large structural change between GTP- and GDP-bound form EF1B (EF-Ts) 30 kDa GEF for EF1A EF2 (EF-G) 77 kDa protein GTP and GDP are bound with similar affinities Therefore, no GEF needed? Higher cellular levels of GTP than GDP Equilbrium towards the GTP-bound form Catalyzes translocation of the ribosome Elongation cycle Elongation cycle Ternary complexes (aa-tRNA?EF1A?GTP) enter ribosome in a “testing phase” where the anticodon is tested for codon matching, aa-tRNA is in A site, EF1A in a Factor-binding site F Many complexes enter, non-cognate - fast offrates, cognate - slow offrates Upon cognate recognition, EF1A?GTP is brought into the GTPase activating center of the ribosome GTP is hydrolyzed EF1A?GDP leaves the ribosome. 3’CCA end of the aa-tRNA enters the A site of the 50S subunit Pe