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Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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Structural basis for tRNA decoding and aminoacylation sensing by T-box riboregulators.

Robert A Battaglia1, Jason C Grigg2, Ailong Ke3

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Nature Structural & Molecular Biology
|November 20, 2019
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T-box riboregulators control bacterial responses to nutrient scarcity. This study reveals the high-resolution structure of a Mycobacterium tuberculosis T-box, explaining how it decodes transfer RNAs (tRNAs) and senses their aminoacylation status.

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Bacterial Genetics

Background:

  • T-box riboregulators are essential cis-regulatory RNAs controlling bacterial gene expression.
  • They respond to amino acid starvation by sensing the aminoacylation status of specific transfer RNAs (tRNAs).
  • Understanding their structure is key to elucidating their regulatory mechanism.

Purpose of the Study:

  • To determine the high-resolution crystal structure of a full-length T-box riboregulator from Mycobacterium tuberculosis.
  • To elucidate the structural basis for tRNA decoding and aminoacylation sensing by T-box riboregulators.

Main Methods:

  • X-ray crystallography
  • High-resolution structural analysis
  • Biophysical characterization (implied)

Main Results:

  • The crystal structure reveals a T-box composed of distinct decoding and aminoacylation sensing modules linked by a pseudoknot.
  • The decoding module involves Stem-I and the Stem-II S-turn, forming a claw-like structure.
  • The aminoacylation sensing module, comprising the antiterminator, Stem-III, and linker, interacts with uncharged tRNA via specific contacts.

Conclusions:

  • The structure provides a detailed mechanistic explanation for how T-box riboregulators recognize and discriminate between charged and uncharged tRNAs.
  • The interaction of the linker region with uncharged tRNA is crucial for sensing aminoacylation status.
  • Structural changes upon binding of charged tRNA mimics suggest a mechanism for altered gene expression output.