Steps in Translation

Initiation à Elongation à Termination

Initiation
• Assembly of the components of translation
• Two ribosomal subunits
• mRNA
• Before initiation, aminoacyl-tRNA synthetases
attach amino acids to their respective tRNAs
(charging)
• Requires hydrolysis of ATP à AMP
• Proofreading is possible to remove incorrect amino
acids
• Initiation factors (IF-1, IF-2, IF-3, eIF)
• GTP (and ATP for eukaryotes)
Initiation
• In eukaryotes, the small (40s) ribosomal subunit
binds close to the cap structure at the 5’-end of the
mRNA and moves 5’ à 3’ along the mRNA until it
encounters the initiation codon (AUG).

• Initiation codon is recognized by a special initiator

tRNA = attachment of Methionine (eukaryotes)/
N-formylated Methionine (prokaryotes)
Initiation
• In both prokaryotic and eukaryotic cells,
this N-terminal Met is usually removed
before translation is completed
Elongation
• A cyclic process on the ribosome in which one
amino acid at a time is added to the growing
peptide chain
• Requires elongation factors
• Steps:
1. Binding of aminoacyl tRNA to the A site
2. Peptide bond formation catalyzed by peptidyl
transferase
3. Translocation of the ribosome on the mRNA
4. Expulsion of the deacylated tRNA from the P- and
E-sites
Termination

• Occurs when one of the three termination

codons moves into the A site

• Results in the release of the newly

synthesized protein and dissociation of
the ribosomes and mRNA
Keypoints in Protein Synthesis

tRNA Wobble Hypothesis

• In most cases, accurate base pairing is
required only in the first two nucleotide
positions of an mRNA codon
• Allows a single tRNA to recognize more
than one codon
Keypoints in Protein Synthesis
Energy Requirement
• Addition of a single amino acid to the
polypeptide chain requires cleavage of four
high-energy bonds from ATP and GTP:
• tRNA aminoacylation: ATP à AMP
• Loading tRNA onto ribosome: GTP à GDP
• Translocation: GTP à GDP
Post-translational
Modifications
• Removal of excess amino acids
• Phosphorylation
• Glycosylation
• Hydroxylation
• Proteins that are defective or destined for rapid
turnover are marked for destruction by ubiquitin
and degraded by proteasomes