NusG is an intrinsic transcription termination factor that stimulates motility and coordinates gene expression with NusA

  1. Zachary F Mandell
  2. Reid T Oshiro
  3. Alexander V Yakhnin
  4. Rishi Vishwakarma
  5. Mikhail Kashlev
  6. Daniel B Kearns
  7. Paul Babitzke  Is a corresponding author
  1. The Pennsylvania State University, United States
  2. Indiana University, United States
  3. National Cancer Institute, United States

Abstract

NusA and NusG are transcription factors that stimulate RNA polymerase pausing in Bacillus subtilis. While NusA was known to function as an intrinsic termination factor in B. subtilis, the role of NusG in this process was unknown. To examine the individual and combinatorial roles that NusA and NusG play in intrinsic termination, Term-seq was conducted in wild type, NusA depletion, DnusG, and NusA depletion DnusG strains. We determined that NusG functions as an intrinsic termination factor that works alone and cooperatively with NusA to facilitate termination at 88% of the 1400 identified intrinsic terminators. Our results indicate that NusG stimulates a sequence-specific pause that assists in the completion of suboptimal terminator hairpins with weak terminal A-U and G-U base pairs at the bottom of the stem. Loss of NusA and NusG leads to global misregulation of gene expression and loss of NusG results in flagella and swimming motility defects.

Data availability

RNA-seq data were deposited in GEO under accession number GSE154522. All other data generated or analysed during this study are included in the manuscript and supporting files.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Zachary F Mandell

    Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Reid T Oshiro

    Department of Biology, Indiana University, Bloomington, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Alexander V Yakhnin

    NCI RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7313-7054
  4. Rishi Vishwakarma

    Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Mikhail Kashlev

    NCI RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Daniel B Kearns

    Department of Biology, Indiana University, Bloomington, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Paul Babitzke

    Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, United States
    For correspondence
    pxb28@psu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2481-1062

Funding

National Institutes of Health (GM098399)

  • Paul Babitzke

National Institutes of Health (GM131783)

  • Daniel B Kearns

National Institutes of Health (intramural)

  • Mikhail Kashlev

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Joseph T Wade, Wadsworth Center, New York State Department of Health, United States

Publication history

  1. Received: August 7, 2020
  2. Accepted: April 8, 2021
  3. Accepted Manuscript published: April 9, 2021 (version 1)
  4. Version of Record published: April 21, 2021 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

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  1. Zachary F Mandell
  2. Reid T Oshiro
  3. Alexander V Yakhnin
  4. Rishi Vishwakarma
  5. Mikhail Kashlev
  6. Daniel B Kearns
  7. Paul Babitzke
(2021)
NusG is an intrinsic transcription termination factor that stimulates motility and coordinates gene expression with NusA
eLife 10:e61880.
https://doi.org/10.7554/eLife.61880

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