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Title: Structural basis of non-canonical transcriptional regulation by the σA-bound iron-sulfur protein WhiB1 in M. tuberculosis

Abstract

Abstract WhiB1 is a monomeric iron–sulfur cluster-containing transcription factor in the WhiB-like family that is widely distributed in actinobacteria including the notoriously persistent pathogen Mycobacterium tuberculosis (M. tuberculosis). WhiB1 plays multiple roles in regulating cell growth and responding to nitric oxide stress in M. tuberculosis, but its underlying mechanism is unclear. Here we report a 1.85 Å-resolution crystal structure of the [4Fe–4S] cluster-bound (holo-) WhiB1 in complex with the C-terminal domain of the σ70-family primary sigma factor σA of M. tuberculosis containing the conserved region 4 (σA4). Region 4 of the σ70-family primary sigma factors is commonly used by transcription factors for gene activation, and holo-WhiB1 has been proposed to activate gene expression via binding to σA4. The complex structure, however, unexpectedly reveals that the interaction between WhiB1 and σA4 is dominated by hydrophobic residues in the [4Fe–4S] cluster binding pocket, distinct from previously characterized canonical σ704-bound transcription activators. Furthermore, we show that holo-WhiB1 represses transcription by interaction with σA4in vitro and that WhiB1 must interact with σA4 to perform its essential role in supporting cell growth in vivo. Together, these results demonstrate that holo-WhiB1 regulates gene expression by a non-canonical mechanism relative to well-characterized σA4-dependent transcription activators.

Authors:
 [1];  [1];  [1];  [1];  [1];  [2];  [3]
  1. Department of Biochemistry
  2. Department of Biochemistry, Redox Biology Center
  3. Department of Biochemistry, Redox Biology Center, Nebraska Center for Integrated Biomolecular Communication, University of Nebraska-Lincoln, Lincoln, NE 68588, USA
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1577377
Grant/Contract Number:  
AC02–76SF00515
Resource Type:
Published Article
Journal Name:
Nucleic Acids Research
Additional Journal Information:
Journal Name: Nucleic Acids Research Journal Volume: 48 Journal Issue: 2; Journal ID: ISSN 0305-1048
Publisher:
Oxford University Press
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Wan, Tao, Li, Shanren, Beltran, Daisy Guiza, Schacht, Andrew, Zhang, Lu, Becker, Donald F., and Zhang, LiMei. Structural basis of non-canonical transcriptional regulation by the σA-bound iron-sulfur protein WhiB1 in M. tuberculosis. United Kingdom: N. p., 2019. Web. doi:10.1093/nar/gkz1133.
Wan, Tao, Li, Shanren, Beltran, Daisy Guiza, Schacht, Andrew, Zhang, Lu, Becker, Donald F., & Zhang, LiMei. Structural basis of non-canonical transcriptional regulation by the σA-bound iron-sulfur protein WhiB1 in M. tuberculosis. United Kingdom. doi:10.1093/nar/gkz1133.
Wan, Tao, Li, Shanren, Beltran, Daisy Guiza, Schacht, Andrew, Zhang, Lu, Becker, Donald F., and Zhang, LiMei. Fri . "Structural basis of non-canonical transcriptional regulation by the σA-bound iron-sulfur protein WhiB1 in M. tuberculosis". United Kingdom. doi:10.1093/nar/gkz1133.
@article{osti_1577377,
title = {Structural basis of non-canonical transcriptional regulation by the σA-bound iron-sulfur protein WhiB1 in M. tuberculosis},
author = {Wan, Tao and Li, Shanren and Beltran, Daisy Guiza and Schacht, Andrew and Zhang, Lu and Becker, Donald F. and Zhang, LiMei},
abstractNote = {Abstract WhiB1 is a monomeric iron–sulfur cluster-containing transcription factor in the WhiB-like family that is widely distributed in actinobacteria including the notoriously persistent pathogen Mycobacterium tuberculosis (M. tuberculosis). WhiB1 plays multiple roles in regulating cell growth and responding to nitric oxide stress in M. tuberculosis, but its underlying mechanism is unclear. Here we report a 1.85 Å-resolution crystal structure of the [4Fe–4S] cluster-bound (holo-) WhiB1 in complex with the C-terminal domain of the σ70-family primary sigma factor σA of M. tuberculosis containing the conserved region 4 (σA4). Region 4 of the σ70-family primary sigma factors is commonly used by transcription factors for gene activation, and holo-WhiB1 has been proposed to activate gene expression via binding to σA4. The complex structure, however, unexpectedly reveals that the interaction between WhiB1 and σA4 is dominated by hydrophobic residues in the [4Fe–4S] cluster binding pocket, distinct from previously characterized canonical σ704-bound transcription activators. Furthermore, we show that holo-WhiB1 represses transcription by interaction with σA4in vitro and that WhiB1 must interact with σA4 to perform its essential role in supporting cell growth in vivo. Together, these results demonstrate that holo-WhiB1 regulates gene expression by a non-canonical mechanism relative to well-characterized σA4-dependent transcription activators.},
doi = {10.1093/nar/gkz1133},
journal = {Nucleic Acids Research},
number = 2,
volume = 48,
place = {United Kingdom},
year = {2019},
month = {12}
}

Journal Article:
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DOI: 10.1093/nar/gkz1133

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