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Structural Basis for NusA Stabilized Transcriptional Pausing.

Xieyang Guo1, Alexander G Myasnikov1, James Chen2

  • 1Department of Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), 67404 Illkirch Cedex, France; Université de Strasbourg, 67404 Illkirch Cedex, France; Centre National de la Recherche Scientifique (CNRS), UMR 7104, 67404 Illkirch Cedex, France; Institut National de la Santé et de la Recherche Médicale (Inserm), U964, 67404 Illkirch Cedex, France.

Molecular Cell
|March 3, 2018
PubMed
Summary
This summary is machine-generated.

The bacterial transcription factor NusA regulates gene expression by stimulating RNA polymerase (RNAP) pausing and termination. Structural studies reveal how NusA interacts with RNAP to control RNA folding and inhibit catalysis during transcription.

Keywords:
NusARNA polymerase structurecryo-EMhis-pausetranscriptiontranscriptional pausing

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Genetics

Background:

  • Transcriptional pausing by RNA polymerases (RNAPs) is crucial for regulating gene expression and transcription termination across all life forms.
  • The bacterial transcription factor NusA is essential for stimulating both pausing and termination, highlighting its central regulatory role.

Purpose of the Study:

  • To elucidate the structural mechanisms by which NusA interacts with paused Escherichia coli RNAP elongation complexes.
  • To understand how NusA stimulates RNA folding, pausing, and termination of transcription.

Main Methods:

  • Single-particle electron cryo-microscopy (cryo-EM) reconstructions.
  • Analysis of NusA bound to paused E. coli RNAP elongation complexes, with and without pause-enhancing RNA hairpin structures.

Main Results:

  • Four key interactions between NusA and RNAP were identified, explaining NusA's role in stimulating RNA folding, pausing, and termination.
  • A structural basis for catalysis inhibition was revealed through an asymmetric translocation intermediate of RNA and DNA, rendering the RNAP active site inactive.
  • Dynamic comparison of RNAP at different pausing stages provided insights into the process and NusA's regulatory function.

Conclusions:

  • NusA plays a critical role in regulating bacterial gene expression by modulating RNAP activity during pausing and termination.
  • The reported structures provide a detailed molecular understanding of NusA-RNAP interactions and their functional consequences.
  • This work offers insights into the dynamic regulation of transcription and the mechanism of transcription termination.