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Ch. 12 - Regulation of Gene Expression in Bacteria and Bacteriophage
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 12, Problem 32b

The function of tRNA synthetases is to attach amino acids to tRNAs. Suppose the tRNA synthetase responsible for attaching tryptophan to tRNA is mutated in a bacterial strain, with the result that the tRNA synthetase functions at about 15% of the efficiency of the wild-type tRNA synthetase. Would formation of the 3–4 stem-loop structure in mRNA be more frequent or less frequent in the mutant strain than in the wild-type strain? Why?

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Understand the role of tRNA synthetases: tRNA synthetases are enzymes that attach specific amino acids to their corresponding tRNAs. This process is critical for accurate translation of mRNA into proteins. In this case, the tRNA synthetase in question is responsible for attaching tryptophan to its corresponding tRNA (tRNA^Trp).
Review the 3–4 stem-loop structure in mRNA: The 3–4 stem-loop structure is part of the attenuation mechanism in bacterial operons, such as the trp operon. When this structure forms, it causes transcription termination, preventing the downstream genes from being transcribed. The formation of this structure depends on the availability of charged tRNA^Trp.
Analyze the impact of the mutation: In the mutant strain, the tRNA synthetase functions at only 15% of the efficiency of the wild-type enzyme. This means that fewer tRNA^Trp molecules will be charged with tryptophan, leading to a lower availability of charged tRNA^Trp in the cell.
Relate the availability of charged tRNA^Trp to the 3–4 stem-loop formation: When charged tRNA^Trp is scarce, ribosomes translating the leader peptide sequence in the trp operon stall at the tryptophan codons. This stalling prevents the formation of the 3–4 stem-loop structure, allowing transcription of the downstream genes to continue. Conversely, if charged tRNA^Trp is abundant, the ribosome does not stall, and the 3–4 stem-loop forms, terminating transcription.
Conclude the effect of the mutation: In the mutant strain, due to the reduced efficiency of the tRNA synthetase, the formation of the 3–4 stem-loop structure would be less frequent compared to the wild-type strain. This is because the scarcity of charged tRNA^Trp increases ribosome stalling, preventing the formation of the termination structure.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

tRNA Synthetases

tRNA synthetases are enzymes that play a crucial role in protein synthesis by attaching specific amino acids to their corresponding tRNA molecules. Each tRNA synthetase is specific to one amino acid and its corresponding tRNA, ensuring that proteins are synthesized accurately according to the genetic code. A mutation that reduces the efficiency of a tRNA synthetase can lead to decreased availability of the amino acid for protein synthesis, potentially affecting the overall translation process.
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Stem-Loop Structures in mRNA

Stem-loop structures in mRNA are formed when complementary sequences within the RNA strand base-pair with each other, creating a loop. These structures can play significant roles in the regulation of gene expression, including the termination of transcription and the modulation of translation. The frequency of stem-loop formation can be influenced by the availability of tRNAs and the efficiency of translation, as they can affect the stability and processing of the mRNA.
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Ribosome Structure

Translation Efficiency

Translation efficiency refers to how effectively ribosomes synthesize proteins from mRNA, which is influenced by the availability of tRNAs and their corresponding amino acids. If a tRNA synthetase is functioning at reduced efficiency, it may lead to a shortage of the associated amino acid, thereby slowing down the translation process. This reduction can impact the formation of secondary structures in mRNA, such as stem-loops, as the ribosome may pause more frequently during translation, allowing more time for these structures to form.
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Translation initiation
Related Practice
Textbook Question

For an E. coli strain with the lac operon genotype I⁺ P⁺ O⁺ Z⁺ Y⁺, identify the level of transcription of the operon genes in each growth medium listed. Specify transcription as 'none,' 'basal,' or 'activated' for each medium, and provide an explanation to justify your answer.

Growth medium contains lactose but no glucose.

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Textbook Question

How could antisense RNA be used as an antibiotic? What types of genes would you target using this scheme?

529
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Textbook Question

The function of tRNA synthetases is to attach amino acids to tRNAs. Suppose the tRNA synthetase responsible for attaching tryptophan to tRNA is mutated in a bacterial strain, with the result that the tRNA synthetase functions at about 15% of the efficiency of the wild-type tRNA synthetase.

How would this mutation affect attenuation of the tryptophan operon? Explain your answer.

467
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Textbook Question

The following hypothetical genotypes have genes A, B, and C corresponding to lacI, lacO, and lacZ, but not necessarily in that order. Data in the table indicate whether β-galactosidase is produced in the presence and absence of the inducer for each genotype. Use these data to identify the correspondence between A, B, and C and the lacI, lacO, and lacZ genes. Carefully explain your reasoning for identifying each gene.

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Textbook Question

Northern blot analysis is performed on cellular mRNA isolated from E. coli. The probe used in the northern blot analysis hybridizes to a portion of the lacY sequence. Below is an example of the gel from northern blot analysis for a wild-type lac⁺ bacterial strain. In this gel, lane 1 is from bacteria grown in a medium containing only glucose (minimal medium). Lane 2 is from bacteria in a medium containing only lactose. Following the style of this diagram, draw the gel appearance for northern blots of the bacteria listed below. In each case, lane 1 is for mRNA isolated after growth in a glucose-containing (minimal) medium, and lane 2 is for mRNA isolated after growth in a lactose-only medium.

lac⁺ bacteria with the genotype I⁺ P⁺ OC Z⁺ Y⁺ 

802
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Textbook Question

Northern blot analysis is performed on cellular mRNA isolated from E. coli. The probe used in the northern blot analysis hybridizes to a portion of the lacY sequence. Below is an example of the gel from northern blot analysis for a wild-type lac⁺ bacterial strain. In this gel, lane 1 is from bacteria grown in a medium containing only glucose (minimal medium). Lane 2 is from bacteria in a medium containing only lactose. Following the style of this diagram, draw the gel appearance for northern blots of the bacteria listed below. In each case, lane 1 is for mRNA isolated after growth in a glucose-containing (minimal) medium, and lane 2 is for mRNA isolated after growth in a lactose-only medium.

lac⁻ bacteria with the genotype I⁺ P⁺ O⁺ Z⁻ Y⁺ 

568
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