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Question:

A series of experiments is being conducted to determine the structure and function of different types of bacterial RNA.  Cultures of Staphylococcus aureus are exposed to chemicals that lyse the bacterial cells, and the RNA molecules are then extracted.  A specific RNA consisting of 90 nucleotides is purified for further analysis.  It is found to contain high amounts of chemically modified bases such as dihydrouridine, pseudouridine, and ribothymidine, and its secondary structure arises from base pairing within the chain.  Which of the following is the most likely composition of the 3'-end of this molecule?

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Explanation:

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Transfer RNA (tRNA) is a form of non-coding RNA composed of 74-93 nucleotides.  Specific tRNAs transfer certain amino acid residues to the growing polypeptide during translation.  tRNA functions by recognizing the 3 base codon on the mRNA being translated through its anticodon region, which contains complementary bases.  The secondary structure of tRNA resembles a cloverleaf and contains the following regions:

  • The acceptor stem is created through the base pairing of the 5'-terminal nucleotides with the 3'-terminal nucleotides.  The CCA tail hangs off the 3' end, with the amino acid bound to the 3' terminal hydroxyl group.  tRNA is "loaded" with the appropriate amino acid by aminoacyl tRNA synthetase.  The acceptor stem helps mediate correct tRNA recognition by the proper aminoacyl tRNA synthetase.

  • A 3' CCA tail is added to the 3' end of tRNA as a posttranscriptional modification in eukaryotes and most prokaryotes.  Several enzymes utilize this tail to help recognize tRNA molecules.

  • The D loop contains numerous dihydrouridine residues, which are modified bases often present in tRNA.  The D loop (along with the acceptor stem and anticodon loop) facilitates correct tRNA recognition by the proper aminoacyl tRNA synthetase.

  • The anticodon loop contains sequences that are complementary to the mRNA codon.  During translation, the ribosome complex selects the proper tRNA based solely on its anticodon sequence.

  • The T loop contains the TΨC sequence that is necessary for binding of tRNA to ribosomes.  The TΨC sequence refers to the presence of ribothymidine, pseudouridine, and cytidine residues.

(Choices A and F)  AUG and UAG are mRNA start and stop codons that initiate and terminate translation, respectively.

(Choices C and D)  After transcription, eukaryotic pre-mRNA undergoes posttranscriptional modification, which includes the addition of a poly-A tail at the 3' end and methylguanosine cap at the 5' end, and the removal of introns.

(Choice E)  A TATA box is an upstream promoter region associated with some genes in eukaryotic organisms.  TATA binding protein binds to this promoter during transcription, unwinding the DNA and initiating separation of the strands.

Educational objective:
Transfer RNA (tRNA)  is a small, noncoding form of RNA that contains chemically modified bases (eg, dihydrouridine, ribothymidine, pseudouridine).  tRNA has a CCA sequence at its 3'-end that is used as a recognition sequence by proteins.  The 3' terminal hydroxyl group of the CCA tail serves as the amino acid binding site.