The elongation cycle is highly conserved across all kingdoms of life. Each cycle of elongation adds one amino acid to the C-terminus of the newly synthesized peptide (Yu et al., 2014). Figure 9 describes the steps involved in the elongation cycle.
Figure 9. Bacterial elongation cycle. Elongation cycle involves sequential addition of amino acid to the growing peptide chain. Aminoacylated tRNA in complex with EF-Tu and GTP interacts with the A-site in the decoding center where the correctness of the codon-anticodon is determined. On accommodation aa-tRNA moves into the PTC after which the ribosome forms the hybrid A/P and P/E state in preparation of translocation. After GTP hydrolysis, dissociation of E-F-Tu-GDP and translocation, new aa-tRNA ternary complex reads the codon on the mRNA and continues this cycle(Voorhees e Ramakrishnan, 2013)
Aminoacylation and Delivery of aa-tRNA to the A-site:
E.coli has 20 amino acids, 20 aminoacyl tRNA synthetases (aaRS) and 41 tRNAs with different anticodons (Berman et al., 2000). The tRNAs are charged with specific amino acids based on their anticodon triplet by the aminoacyltRNA synthetase (aaRS). In an ATP-dependent reaction the amino acid is transferred to the 2’ or 3’ hydroxyl group of terminal adenine (A76) at the 3’CCA terminus as shown in figure 10. This process employs dual discrimination based on the rate of reaction and substrate binding affinity (Reynolds et al., 2010) Figure 12. After aminoacylation, the charged tRNA forms a ternary complex with EF-Tu and GTP and is ready to be delivered to the A-site of the ribosome (Shepherd e Ibba, 2015).
Figure 9. Formation of an aminoacylated-tRNA molecule. aaRS activates an amino acid in presence of an ATP mole...
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...he termination phase of protein synthesis is reached. Stop codons are recognized by RF1 or RF2 position the universally conserved GGQ motif into the peptidyl transferase center and catalyze the hydrolysis of peptidyl –tRNA (Wilson, 2014).
Figure 15. Translocation dynamics. 30S head is … in color, 30S body is … in color, 50S subunit is in … color and EF-G is in .. color. a) Pretranslocation state. The tRNA anticodons are located in the A and the P site on the SSU while the tRNA CCA ends oscillate between A and P or P and E site on the 50S subunit. b) Hybrid state. Racheting of SSU head brings tRNa anticodons and mRNA codons into a hybrid stat between A and P(A/P) or P and E (P/E) on SSU. c) Posttranslocation. One tRNA is bound to the ribosome in the P-site and one is bound in the E-site. SSU head is rotated backward and normal position is restored (Rodnina, 2013).
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