Translation: Protein Synthesis

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Translation: Components - The Protein Factory Crew

  • Ribosomes: Site of protein synthesis.
    • Prokaryotes: 70S (50S+30S); Eukaryotes: 80S (60S+40S).
    • Sites: A (aminoacyl), P (peptidyl), E (exit).
  • mRNA: Carries genetic code as codons (AUG start; UAA, UAG, UGA stop).
  • tRNA: Anticodon pairs with mRNA codon; 3'-CCA attaches specific amino acid.
  • Aminoacyl-tRNA Synthetases: "Charge" tRNAs; crucial proofreading.

Ribosome APE sites with mRNA, tRNA, polypeptide

⭐ Ribosomal RNA (rRNA) in the large subunit (e.g., 23S in prokaryotes, 28S in eukaryotes) possesses peptidyl transferase activity, making it a ribozyme.

Translation: Initiation - Ready, Set, Synthesize!

Prokaryotic Initiation (forms 70S complex):

  • mRNA's Shine-Dalgarno sequence (📌 S-D for Small ribosome) aligns with 16S rRNA (30S subunit).
  • Initiation Factors: IF1, IF2-GTP (binds fMet-tRNA), IF3.
  • Initiator tRNA: fMet-tRNA directly to P-site.

Eukaryotic Initiation (forms 80S complex):

  • 40S subunit (with Met-tRNA, eIF2-GTP) binds mRNA's 5' cap, scans to Kozak sequence (📌 Kozak for euKaryotes) around AUG.
  • Initiation Factors: Multiple eIFs (e.g., eIF4E binds cap).

⭐ In prokaryotes, the Shine-Dalgarno sequence on mRNA base-pairs with a complementary sequence on the 16S rRNA of the 30S ribosomal subunit to position it correctly for initiation.

Translation: Elongation - Building the Chain

  • Polypeptide chain grows via a cycle. 📌 APE sites: Aminoacyl, Peptidyl, Exit.
  • Elongation Cycle (GTP-powered):
    • 1. Codon Recognition: aa-tRNA + EF-Tu/eEF1A enters A-site.
    • 2. Peptide Bond Formation: Peptidyl transferase (rRNA) forms peptide bond; chain moves to A-site tRNA.
    • 3. Translocation: EF-G/eEF2 moves ribosome; tRNAs shift (A→P, P→E & exit).

⭐ EF-Tu (prokaryotes) or eEF1A (eukaryotes) is responsible for bringing the charged aminoacyl-tRNA to the A site of the ribosome, a step requiring GTP hydrolysis.

Protein Synthesis Elongation Steps

Translation: Termination - It's a Wrap!

  • Stop Codons: UAA, UAG, UGA. 📌 (U Are Away, U Go Away, U Are Gone).
  • Release Factors (RFs):
    • Prokaryotes: RF1, RF2, RF3.
    • Eukaryotes: eRF1, eRF3.
  • Mechanism:
    • RFs bind stop codon at A-site.
    • Polypeptide hydrolysis from tRNA.
    • Ribosomal subunits, mRNA, tRNA dissociate.

⭐ Stop codons are recognized by protein Release Factors (RFs), not by tRNAs, leading to the termination of translation.

Post-Translational Mods - Protein's Extreme Makeover

  • Protein Folding: Chaperones (e.g., Hsp70) ensure correct 3D structure.
  • Covalent Modifications:
    • Phosphorylation, glycosylation, ubiquitination.
    • Acetylation, methylation, proteolytic cleavage.
    • Disulfide bond formation.
  • Significance: Critical for protein function, stability, and diversity.

⭐ Glycosylation, the addition of sugar moieties to proteins, typically begins in the endoplasmic reticulum and continues in the Golgi apparatus, crucial for protein folding, stability, and targeting.

Protein Synthesis Inhibitors - Antibiotic Hit List

📌 Mnemonic: "Buy AT 30, CCELL at 50"

AntibioticTarget Subunit(s)MechanismOrganism
Streptomycin30SMisreading mRNA, inhibits initiation (high conc.)Prokaryotic
Tetracyclines30SBlocks A-siteProkaryotic
Chloramphenicol50SBlocks peptidyl transferaseProkaryotic
Macrolides50SBlocks translocationProkaryotic
Clindamycin50SBlocks translocationProkaryotic
Linezolid50SBlocks initiation complex formationProkaryotic
PuromycinA-sitePremature terminationBoth
Cycloheximide80S (Euk)Blocks translocationEukaryotic
Diphtheria ToxineEF2Inactivates eEF2 (ADP-ribosylation)Eukaryotic

High‑Yield Points - ⚡ Biggest Takeaways

  • Translation occurs on ribosomes (70S prokaryotes, 80S eukaryotes), decoding mRNA to protein.
  • mRNA codons pair with tRNA anticodons carrying specific amino acids.
  • Initiation: AUG start codon (Met); Shine-Dalgarno (prok) / Kozak sequence (euk).
  • Elongation: Peptidyl transferase (rRNA ribozyme) forms peptide bonds in A, P, E sites.
  • Termination: Stop codons (UAA, UAG, UGA) trigger release of polypeptide.
  • Key inhibitors: Many antibiotics (tetracyclines, macrolides) target 70S ribosomes.

Practice Questions: Translation: Protein Synthesis

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Which of the following statements regarding collagen synthesis is incorrect?

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Flashcards: Translation: Protein Synthesis

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Glutamine-PRPP amidotransferase (purine synthesis) is regulated by negative feedback via _____, IMP, and GMP

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Glutamine-PRPP amidotransferase (purine synthesis) is regulated by negative feedback via _____, IMP, and GMP

AMP

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