Question

Describe and give the results of an experiment that shows that polymerase III makes tRNA and $5 \mathrm{~S}$ rRNA.

   Describe and give the results of an experiment that shows that polymerase III makes tRNA and $5 \mathrm{~S}$ rRNA.
Molecular Biology
Molecular Biology
Robert F. Weaver 5th Edition
Chapter 10, Problem 3 ↓

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To do this, we need to selectively inhibit or knock out polymerase III activity in a cell and observe the effects on the synthesis of tRNA and 5S rRNA.  Show more…

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Describe and give the results of an experiment that shows that polymerase III makes tRNA and $5 \mathrm{~S}$ rRNA.
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Key Concepts

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5S rRNA
5S ribosomal RNA (rRNA) is a component of the ribosome that plays a structural and functional role in protein synthesis. In eukaryotes, its transcription by RNA Polymerase III is a clear example of the enzyme's specialized function in producing non-coding RNAs vital for ribosome assembly.
tRNA
Transfer RNA (tRNA) is a small RNA molecule required for translating mRNA codons into amino acids during protein synthesis. Its synthesis by RNA Polymerase III underscores the enzyme's role in creating essential components of the protein translation machinery.
Experimental Approaches in Transcription Studies
Experimental methods such as the use of specific inhibitors (e.g., alpha-amanitin at concentrations that selectively inhibit RNA Polymerase II) and transcription run-on assays are critical in distinguishing the activities of different RNA polymerases. These approaches help to demonstrate that RNA Polymerase III is uniquely responsible for synthesizing tRNA and 5S rRNA by finding that these transcripts are produced even when other polymerases are inhibited.
RNA Polymerases
RNA polymerases are essential enzymes that synthesize RNA molecules using a DNA template. In eukaryotic cells, different RNA polymerases (I, II, III) are specialized to transcribe specific classes of RNA, such as rRNA, mRNA, and small structural RNAs. Understanding their roles is fundamental in studying gene expression and regulation.
RNA Polymerase III
RNA Polymerase III is responsible for transcribing small, non-coding RNAs including tRNA and 5S rRNA. This enzyme operates with distinct regulatory mechanisms and inhibitor sensitivities, making it possible to identify its activity in experiments that differentiate between the products of various RNA polymerases.

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2. Investigators have discovered that yeast cells synthesize an RNA molecule that is not mRNA, rRNA, tRNA, or any other known RNA species. They wish to know whether RNA polymerase I, II, or III is responsible for its synthesis. Suggest a simple experimental approach that would help to determine which of the polymerases is responsible for the synthesis of the new type of RNA. 3. In contrast to bacteria, eukaryotes require a distinct RNA polymerase to synthesize rRNAs, mRNAs, and tRNAs. Is this requirement for a distinct RNA polymerase a result of the fact that eukaryotes have a defined nucleus and bacteria do not? Provide evidence to support your answer.

scientists-studying-transcription-in-yeast-saccharomyces-cerevisiae-created-an-experimental-strain-that-produced-a-modified-rna-polymerase-containing-a-single-amino-acid-substitution-the-sci-83683

Scientists studying transcription in yeast (Saccharomyces cerevisiae) created an experimental strain that produced a modified RNA polymerase containing a single amino acid substitution. The scientists determined the maximum elongation rate during transcription with and without the modified RNA polymerase enzyme (Figure 1). The compound amanitin, which is commonly found in toxic mushrooms, is a specific RNA polymerase inhibitor. Amanitin binds to the RNA polymerase active site and inhibits transcription. In a second experiment, the scientists treated the wild-type and experimental strains of S. cerevisiae with a 40 μg/mL solution of amanitin and recorded the maximum elongation rate of the mRNA (Figure 2). Error bars represent ±2SEx. Figure 1. Maximum elongation rate under natural conditions Figure 2. Maximum elongation rate in a 40 μg/mL amanitin solution

scientists-studying-transcription-in-yeast-saccharomyces-cerevisiae-created-an-experimental-strain-that-produced-modified-rna-polymerase-containing-single-amino-acid-substitution-the-scienti-72598

Scientists studying transcription in yeast (Saccharomyces cerevisiae) created an experimental strain that produced a modified RNA polymerase containing a single amino acid substitution. The scientists determined the maximum elongation rate during transcription with and without the modified RNA polymerase enzyme (Figure 1). The compound amanitin, which is commonly found in toxic mushrooms, is a specific RNA polymerase inhibitor. Amanitin binds to the RNA polymerase active site and inhibits transcription. In a second experiment, the scientists treated the wild-type and experimental strains of S. cerevisiae with a 40 μg/mL solution of amanitin and recorded the maximum elongation rate of the mRNA (Figure 2). Error bars represent ±2SEx̄. Figure 1. Maximum elongation rate under natural conditions Figure 2. Maximum elongation rate in a 40 μg/mL amanitin solution

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