Q.(a) Explain the process by which amino acid gets attached to the tRNA molecule during translation process.
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Start your 14-day free trial to unlock the full solution →Aminoacyl-tRNA synthetases catalyze the attachment of amino acids to their cognate tRNAs in a two-step ATP-dependent reaction. Translation terminates when a stop codon is recognized by release factors, triggering peptide release. UTR stands for Untranslated Region, found at the 5′ and 3′ ends of mRNA flanking the coding sequence.
(a) Aminoacylation: Charging the tRNA
The attachment of an amino acid to its corresponding tRNA is called aminoacylation or tRNA charging. This reaction is catalyzed by a family of enzymes called aminoacyl-tRNA synthetases, and it's absolutely critical because it establishes the genetic code—each tRNA must carry the correct amino acid that matches its anticodon.
The process unfolds in two distinct steps, both occurring in the active site of the synthetase:
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Activation of the amino acid
The enzyme first binds both the amino acid and ATP. It catalyzes the formation of an aminoacyl-adenylate intermediate:
Amino acid + ATP → Aminoacyl-AMP + PPi
The amino acid is now "activated" by being linked to AMP through a high-energy bond. The release of pyrophosphate (PPi) makes this step irreversible when the pyrophosphate is hydrolyzed.
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Transfer to tRNA
The activated aminoacyl-AMP remains bound to the enzyme. The cognate tRNA (the one with the correct anticodon) now binds, and the enzyme transfers the amino acid to the 3′ end of the tRNA, specifically to the hydroxyl group of the terminal adenosine nucleotide:
Aminoacyl-AMP + tRNA → Aminoacyl-tRNA + AMP
The amino acid is attached via an ester bond to either the 2′-OH or 3′-OH of the ribose (it can migrate between the two).
The charged aminoacyl-tRNA is now ready to participate in translation. Each of the 20 amino acids has at least one specific aminoacyl-tRNA synthetase that recognizes both the amino acid and the set of tRNAs corresponding to it—this is the molecular basis of the genetic code's fidelity.
The specificity of aminoacyl-tRNA synthetases is so high that the error rate is only about 1 in 10,000. Some synthetases even have "proofreading" sites that hydrolyze incorrectly attached amino acids.
(b) Termination of Translation
Translation doesn't simply run out of mRNA and stop—it requires a specific signal and molecular machinery to release the completed polypeptide.
The signal: Translation terminates when the ribosome encounters one of three stop codons (also called nonsense or termination codons) in the mRNA: UAA, UAG, or UGA. These codons do not code for any amino acid.
The mechanism:
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Recognition by release factors
When a stop codon enters the A site of the ribosome, no aminoacyl-tRNA can bind to it (because no tRNA has an anticodon for these codons). Instead, proteins called release factors (RFs) recognize and bind to the stop codon.
- In prokaryotes: RF1 recognizes UAA and UAG; RF2 recognizes UAA and UGA.
- In eukaryotes: eRF1 recognizes all three stop codons.
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Hydrolysis of the peptidyl-tRNA bond
The release factor triggers the peptidyl transferase center of the ribosome to catalyze hydrolysis instead of peptide bond formation. Water attacks the ester bond linking the polypeptide chain to the tRNA in the P site:
Peptidyl-tRNA + H2O → Polypeptide + tRNA
The completed polypeptide is released from the ribosome.
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Ribosome disassembly
A second release factor (RF3 in prokaryotes, eRF3 in eukaryotes) binds and, using GTP hydrolysis, helps dissociate the ribosomal subunits from the mRNA. The ribosome is now recycled for another round of translation.
Premature stop codons arising from mutations (nonsense mutations) lead to truncated, usually nonfunctional proteins—a common cause of genetic diseases.
(c) UTR: Untranslated Regions …
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