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Molecular Biology Lecture 13
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Molecular Biology Lecture 13

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Questions and Answers

What is a key difference between DNA replication and transcription in terms of chain growth?

  • DNA replication and transcription both involve rapid chain growth at many sites.
  • DNA replication involves slower growth at few intracellular sites, while transcription involves rapid chain growth at many sites.
  • DNA replication involves rapid chain growth at few intracellular sites, while transcription involves slower growth at many sites. (correct)
  • DNA replication involves slower growth at many sites, while transcription involves rapid chain growth at few intracellular sites.
  • What is a characteristic of RNA polymerase subunits in the three domains of life?

  • Homologous subunits are not color coded.
  • Subunits are arranged by size rather than function.
  • Subunits are arranged by function rather than size. (correct)
  • Subunits marked with an asterisk are not conserved among the three eukaryotic RNA polymerases.
  • What is shown in the crystal structures of RNA polymerases?

  • Core enzyme structures with bound proteins.
  • Bound proteins that aid in template recognition.
  • Core enzyme structures without bound proteins. (correct)
  • Only the promoter site of the DNA.
  • What is the first step in the transcription process by bacterial RNA polymerase?

    <p>Sequence-specific interaction between RNA polymerase and a promoter site.</p> Signup and view all the answers

    What is a feature of the open-promoter complex formed by Taq RNA polymerase?

    <p>A sharp bend in the template strand.</p> Signup and view all the answers

    What is the function of the RNA polymerase subunits marked with an asterisk?

    <p>They are conserved among the three eukaryotic RNA polymerases.</p> Signup and view all the answers

    What is a difference between DNA replication and transcription in terms of RNA accumulation?

    <p>More RNA accumulates than DNA.</p> Signup and view all the answers

    What is the characteristic of the promoter site in transcription?

    <p>It is where transcription begins.</p> Signup and view all the answers

    What is the consequence of backtracking in an elongation complex?

    <p>The transcript terminus is cleaved, creating a new base-paired 3' terminus.</p> Signup and view all the answers

    What determines the frequency of transcription of E. coli genes?

    <p>The similarity between the gene's promoter sequence and the consensus sequence.</p> Signup and view all the answers

    What is the function of the protein in factor-dependent termination?

    <p>It acts as an RNA-DNA helicase.</p> Signup and view all the answers

    What is the composition of the GC-rich region in factor-independent termination?

    <p>A region with high GC content.</p> Signup and view all the answers

    What happens to the RNA-RNA duplex in factor-independent termination?

    <p>It is stabilized by G-C base pairs.</p> Signup and view all the answers

    What is the consequence of the RNA-RNA duplex formation in factor-independent termination?

    <p>The transcription is terminated.</p> Signup and view all the answers

    What is the role of the A-rich segment in factor-independent termination?

    <p>It is transcribed into a U-rich mRNA segment.</p> Signup and view all the answers

    What is the purpose of the run of four to eight A residues in factor-independent termination?

    <p>To facilitate release of the transcript.</p> Signup and view all the answers

    What is the purpose of phosphorylation in transcription initiation?

    <p>To release the enzyme from the initiation site</p> Signup and view all the answers

    What is the function of the AATAAA sequence near the end of most eukaryotic genes?

    <p>It provides a signal for endonuclease cleavage and poly(A) tail addition</p> Signup and view all the answers

    What is the location of acetylation in core histones?

    <p>In the N-terminal domain</p> Signup and view all the answers

    What is the purpose of DNA looping in transcriptional regulation?

    <p>To bring enhancer-bound activator proteins into contact with TAFs associated with the core complex</p> Signup and view all the answers

    Which of the following is NOT a step in post-transcriptional processing in bacteria?

    <p>Splicing of exons</p> Signup and view all the answers

    What is the function of the TFIIA–TBP–TFIIB complex?

    <p>To initiate transcription by binding to the promoter</p> Signup and view all the answers

    Which of the following is a step in post-transcriptional processing in eukaryotes?

    <p>Addition of a poly(A) tail</p> Signup and view all the answers

    What is the structure of the core histones?

    <p>A helical domain plus a highly basic N-terminal domain</p> Signup and view all the answers

    What is the primary function of alanyl-tRNA synthetase?

    <p>To aminoacylate tRNA</p> Signup and view all the answers

    What is the characteristic of the cleft in the glutaminyl-tRNA synthetase?

    <p>It accommodates both ATP and the amino acid</p> Signup and view all the answers

    What is the difference between bacterial and eukaryotic ribosomes?

    <p>Size of the ribosomal subunits</p> Signup and view all the answers

    What is the method used to determine the shape of the ribosomal subunits?

    <p>Electron microscopy</p> Signup and view all the answers

    What is the characteristic of the assembly pathway of the 30S subunit?

    <p>It is an obligatory process</p> Signup and view all the answers

    What is the function of the 3' acceptor stem of the tRNA?

    <p>To accommodate the amino acid</p> Signup and view all the answers

    What is the class of the glutaminyl-tRNA synthetase?

    <p>Monomeric class I</p> Signup and view all the answers

    What is the component of the tRNA that is represented by a detailed atomic model?

    <p>The entire tRNA molecule</p> Signup and view all the answers

    What is the function of SF3b in post-transcriptional processing?

    <p>To act as a protein splicing factor</p> Signup and view all the answers

    What is the significance of alternative splicing in gene expression?

    <p>It allows one gene to specify several proteins</p> Signup and view all the answers

    What is the function of NTC in post-transcriptional processing?

    <p>To act as a multiprotein complex called Prp19</p> Signup and view all the answers

    What is the purpose of the smooth (SM) and striated (STR) exons?

    <p>To encode amino acid residues 39-80</p> Signup and view all the answers

    What is the process by which the ribosome selects specific aminoacyl-tRNAs during translation?

    <p>By matching the trinucleotide anticodon on the tRNA to the trinucleotide codon on the RNA message</p> Signup and view all the answers

    What is the result of alternative 3'-terminal exons in the tropomyosin gene?

    <p>Production of different mRNAs with the same coding sequence</p> Signup and view all the answers

    What is the term for the process of selecting specific splice sites during alternative splicing?

    <p>Splice-site selection</p> Signup and view all the answers

    What is the outcome of the tropomyosin gene organization?

    <p>Production of multiple proteins with different functions</p> Signup and view all the answers

    Study Notes

    Enzymology of RNA Synthesis: RNA Polymerase

    • RNA polymerase is responsible for transcription, which involves slower growth at many sites, resulting in more RNA accumulation than DNA
    • The functions of RNA polymerase subunits can be determined by reconstitution
    • RNA polymerase subunit structures vary in the three domains of life (bacteria, archaea, and eukarya), with homologous subunits color-coded and conserved subunits marked with an asterisk

    Mechanism of Transcription

    • Transcription begins with sequence-specific interaction between RNA polymerase and a promoter site, involving duplex unwinding and template strand selection
    • DNA bending occurs in the open-promoter complex, with a sharp bend in the template strand
    • Backtracking can occur in the elongation complex, where the enzyme slips backward, leaving the transcript terminus at the end of a non-base-paired RNA tail
    • Transcription can resume either by forward sliding of the polymerase back to the original structure or by cleavage of the non-base-paired part of the transcript, creating a new base-paired 3' terminus
    • The frequency of gene transcription in E. coli is largely determined by the similarity between the gene's promoter sequence and the consensus sequence
    • Conserved sequences in promoters recognized by E. coli RNA polymerase include the -10 and -35 regions, with lengths of spacer sequences also shown

    Mechanism of Transcription (continued)

    • DNA sequences that promote factor-independent termination include a run of four to eight A residues and a GC-rich region that forms a stem-loop
    • In factor-dependent termination, a protein acts as an RNA-DNA helicase, unwinding the template-transcript duplex and facilitating release of the transcript
    • A model for factor-independent termination of transcription involves an A-rich segment of the template being transcribed into a U-rich mRNA segment, followed by RNA-RNA duplex formation and elimination of some base pairing between the template and transcript

    Transcription and its Control in Eukaryotic Cells

    • A model of the TFIIA-TBP-TFIIB-promoter complex based on crystal structures of TBP-TFIIB-TATA and TFIIA-TBP-TATA complexes
    • DNA looping (perhaps mediated by nucleosomes) can bring enhancer-bound activator (or repressor) proteins into contact with TAFs associated with the core complex
    • Acetylation of core histones occurs exclusively in the N-terminal domains, at highly conserved sites

    Termination of Transcription in Eukaryotes

    • Termination of transcription in eukaryotes involves the addition of poly(A) tails, which is signaled by the AATAAA sequence near the end of most eukaryotic genes

    Post-transcriptional Processing

    • Bacterial transcripts undergo post-transcriptional processing, involving both endonucleolytic and exonucleolytic cleavage
    • Post-transcriptional processing in bacteria involves cleavage of the primary transcript, modification of bases, non-transcriptive nucleotide addition, and intron splicing (in a few cases)
    • Alternative splicing allows one gene to specify several proteins, as seen in the tropomyosin gene organization (rat) and its seven alternative splicing pathways

    Post-transcriptional Processing (continued)

    • Possible mechanisms for alternative splicing by splice-site selection include the use of different splice sites, the presence of splicing enhancers or silencers, and the role of RNA-binding proteins

    Translation

    • Translation of an RNA message into a protein involves the ribosome moving along the message, accepting specific aminoacyl-tRNAs in succession, and selecting them by matching the trinucleotide anticodon on the tRNA to the trinucleotide codon on the RNA message

    The Major Participants in Translation

    • Major "identity elements" in tRNAs include the acceptor stem, the D stem, the anticodon stem, and the variable loop
    • A model of the E. coli glutaminyl-tRNA synthetase coupled with its tRNA and ATP shows the tRNA fitting into a deep cleft in the synthetase, with the ATP and 3' acceptor stem of the tRNA binding to the synthetase
    • Components of bacterial and eukaryotic ribosomes include the 30S and 50S subunits, with eukaryotic ribosomes being somewhat larger and more complex

    The Major Participants in Translation (continued)

    • Assembly map for the 30S subunit shows the obligatory nature of some protein-binding events

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    Description

    This quiz covers the transcription and post-transcriptional processing, translation and post-translational protein processing, and enzymology of RNA synthesis.

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