The Steps of Translation 1. Initiation The small subunit of the ribosome binds to a site "upstream" (on the 5' side) of the start of the message. It proceeds downstream (5' -> 3') until it encounters the start codon AUG. (The region between the cap and the AUG is known as the 5'- untranslated region [5'-UTR].) Here it is joined by the large subunit and a special initiator tRNA. The initiator tRNA binds to the P site (shown in pink) on the ribosome. In eukaryotes, initiator tRNA carries methionine (Met). (Bacteria use a modified methionine designated fMet.) 2. Elongation An aminoacyl-tRNA (a tRNA covalently bound to its amino acid) able to base pair with the next codon on the mRNA arrives at the A site (green) associated with: o an elongation factor (called EF-Tu in bacteria) o GTP (the source of the needed energy) The preceding amino acid (Met at the start of translation) is covalently linked to the incoming amino acid with a peptide bond (shown in red). The initiator tRNA is released from the P site. The ribosome moves one codon downstream. This shifts the more recently-arrived tRNA, with its attached peptide, to the P site and opens the A site for the arrival of a new aminoacyl-tRNA. This last step is promoted by another protein elongation factor (called EF-G in bacteria) and the energy of another molecule of GTP. Note: the initiator tRNA is the only member of the tRNA family that can bind directly to the P site. The P site is so-named because, with the exception of initiator tRNA, it binds only to a peptidyl-tRNA molecule; that is, a tRNA with the growing peptide attached. The A site is so-named because it binds only to the incoming aminoacyl-tRNA; that is the tRNA bringing the next amino acid. So, for example, the tRNA that brings Met into the interior of the polypeptide can bind only to the A site. 3. Termination The end of translation occurs when the ribosome reaches one or more STOP codons (UAA, UAG, UGA). (The nucleotides from this point to the poly(A) tail make up the 3'- untranslated region [3'-UTR] of the mRNA.) There are no tRNA molecules with anticodons for STOP codons. (With a few special exceptions: link to mitochondrial genes and to nonstandard amino acids.) However, protein release factors recognize these codons when they arrive at the A site. Binding of these proteins —along with a molecule of GTP — releases the polypeptide from the ribosome. The ribosome splits into its subunits, which can later be reassembled for another round of protein synthesis.