Question 1

What is the role of tRNA in protein synthesis?

Accepted Answer

Transfer RNAs (tRNAs) are non-coding RNAs essential for protein synthesis. Each tRNA carries a specific amino acid and contains an anticodon that recognizes corresponding codons on mRNA. Eukaryotic cells contain over 50 distinct tRNAs, enabling accurate translation of genetic information into proteins.

Question 2

How does RNA Polymerase III transcribe tRNA genes?

Accepted Answer

RNA Polymerase III transcribes tRNA genes into long precursor tRNA (pre-tRNA) molecules. The pre-tRNA contains a 5' leader sequence, a 3' trailer sequence with a polyuridine tract, a 14-nucleotide intron, and unmodified bases. This initial transcript requires extensive post-transcriptional processing before becoming a functional mature tRNA.

Question 3

What are the main steps in tRNA processing?

Accepted Answer

tRNA processing involves five key steps: removal of the 5' leader by RNase P, trimming the 3' trailer by exonucleases like RNase D, addition of the CCA sequence by tRNA nucleotidyltransferase, chemical modification of specific bases, and intron splicing. The extent and order of processing varies among different tRNAs.

Question 4

Why are modified bases important in tRNA structure?

Accepted Answer

Modified bases, which comprise nearly 20% of some tRNAs, protect the molecule from enzymatic degradation by RNases. These modifications include methylation, deamination, reduction, and isomerization, each catalyzed by specific enzymes with unique base and site-specificity. Modified bases also enable proper tRNA recognition and function during translation.

Question 5

How do the three tRNA hairpin loops differ in function?

Accepted Answer

The TΨC arm is recognized by the ribosome during translation. The D arm, containing dihydrouracil, serves as a recognition site for aminoacyl-tRNA synthetase, which catalyzes covalent amino acid attachment. The anticodon loop often contains queuine, a modified guanine that enables wobble base pairing with mRNA codons, improving translation accuracy.

Question 6

What is wobble base pairing and why does it matter?

Accepted Answer

Wobble base pairing occurs when modified bases like queuine in the anticodon form non-Watson-Crick base pairs with mRNA codons. This loose pairing at the third codon position allows one tRNA to recognize multiple codon variants. The presence of queuine improves translation accuracy by enabling proper codon recognition.

Question 7

How do tRNA modifications differ between eukaryotes and prokaryotes?

Accepted Answer

The nature and position of modified bases are species-specific. Thiolation of adenine occurs only in prokaryotes, while methylation of cytosine is restricted to eukaryotes. Overall, eukaryotic tRNAs undergo more extensive modifications than prokaryotic tRNAs, though both organisms heavily modify the three stem-loop regions.

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