Structure and function of the mycobacterial transcription initiation complex with the essential regulator RbpA

  1. Elizabeth A Hubin
  2. Allison Fay
  3. Catherine Xu
  4. James M Bean
  5. Ruth M Saecker
  6. Michael S Glickman
  7. Seth A Darst  Is a corresponding author
  8. Elizabeth A Campbell  Is a corresponding author
  1. The Rockefeller University, United States
  2. Sloan-Kettering Institute, United States
  3. University of Cambridge, United States

Abstract

RbpA and CarD are essential transcription regulators in mycobacteria. Mechanistic analyses of promoter open complex (RPo) formation establish thatRbpA and CarD cooperatively stimulate formation of anintermediate (RP2) leading to RPo; formation of RP2 islikely a bottleneck step at the majority of mycobacterial promoters. Once RPo forms, CarD also disfavors its isomerization back to RP2.We determined a 2.76 Å-resolution crystal structure of a mycobacterial transcription initiation complex (TIC) with RbpA as well as a CarD/RbpA/TIC model. Both CarD and RbpA bind near the upstream edge of the -10 element where they likely facilitate DNA bending and impede transcription bubble collapse. In vivo studies demonstrate the essential role of RbpA, effects of RbpA truncations on transcription and cell physiology, and indicate additional functions for RbpA not evident in vitro. This work provides a framework to understand the control of mycobacterial transcriptionby RbpA and CarD.

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

Article and author information

Author details

  1. Elizabeth A Hubin

    The Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Allison Fay

    Immunology Program, Sloan-Kettering Institute, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Catherine Xu

    Department of Chemistry, University of Cambridge, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. James M Bean

    Immunology Program, Sloan-Kettering Institute, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Ruth M Saecker

    Department of Chemistry, University of Cambridge, Cambridge, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Michael S Glickman

    Immunology Program, Sloan-Kettering Institute, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7918-5164
  7. Seth A Darst

    The Rockefeller University, New York, United States
    For correspondence
    darst@rockefeller.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8241-3153
  8. Elizabeth A Campbell

    The Rockefeller University, New York, United States
    For correspondence
    elizabeth.campbell0@gmail.com
    Competing interests
    The authors declare that no competing interests exist.

Funding

National Institutes of Health (RO1 GM114450)

  • Elizabeth A Campbell

National Institutes of Health (P30 CA008748)

  • Michael S Glickman

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

Reviewing Editor

  1. James M Berger, Johns Hopkins University School of Medicine, United States

Publication history

  1. Received: October 19, 2016
  2. Accepted: January 7, 2017
  3. Accepted Manuscript published: January 9, 2017 (version 1)
  4. Version of Record published: February 10, 2017 (version 2)

Copyright

© 2017, Hubin et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Elizabeth A Hubin
  2. Allison Fay
  3. Catherine Xu
  4. James M Bean
  5. Ruth M Saecker
  6. Michael S Glickman
  7. Seth A Darst
  8. Elizabeth A Campbell
(2017)
Structure and function of the mycobacterial transcription initiation complex with the essential regulator RbpA
eLife 6:e22520.
https://doi.org/10.7554/eLife.22520

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