8E74

Mycobacterium tuberculosis RNAP paused elongation complex with NusG transcription factor


Experimental Data Snapshot

  • Method: ELECTRON MICROSCOPY
  • Resolution: 2.94 Å
  • Aggregation State: PARTICLE 
  • Reconstruction Method: SINGLE PARTICLE 

wwPDB Validation   3D Report Full Report


This is version 1.1 of the entry. See complete history


Literature

Structural and functional basis of the universal transcription factor NusG pro-pausing activity in Mycobacterium tuberculosis.

Delbeau, M.Omollo, E.O.Froom, R.Koh, S.Mooney, R.A.Lilic, M.Brewer, J.J.Rock, J.Darst, S.A.Campbell, E.A.Landick, R.

(2023) Mol Cell 83: 1474-1488.e8

  • DOI: https://doi.org/10.1016/j.molcel.2023.04.007
  • Primary Citation of Related Structures:  
    8E74, 8E79, 8E82, 8E8M, 8E95

  • PubMed Abstract: 

    Transcriptional pauses mediate regulation of RNA biogenesis. DNA-encoded pause signals trigger pausing by stabilizing RNA polymerase (RNAP) swiveling and inhibiting DNA translocation. The N-terminal domain (NGN) of the only universal transcription factor, NusG/Spt5, modulates pausing through contacts to RNAP and DNA. Pro-pausing NusGs enhance pauses, whereas anti-pausing NusGs suppress pauses. Little is known about pausing and NusG in the human pathogen Mycobacterium tuberculosis (Mtb). We report that MtbNusG is pro-pausing. MtbNusG captures paused, swiveled RNAP by contacts to the RNAP protrusion and nontemplate-DNA wedged between the NGN and RNAP gate loop. In contrast, anti-pausing Escherichia coli (Eco) NGN contacts the MtbRNAP gate loop, inhibiting swiveling and pausing. Using CRISPR-mediated genetics, we show that pro-pausing NGN is required for mycobacterial fitness. Our results define an essential function of mycobacterial NusG and the structural basis of pro- versus anti-pausing NusG activity, with broad implications for the function of all NusG orthologs.


  • Organizational Affiliation

    Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA. Electronic address: darst@rockefeller.edu.


Macromolecules

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Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
Transcription termination/antitermination protein NusGA [auth Z]238Mycobacterium tuberculosisMutation(s): 0 
Gene Names: nusG
UniProt
Find proteins for P9WIU9 (Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv))
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Go to UniProtKB:  P9WIU9
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UniProt GroupP9WIU9
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Entity ID: 2
MoleculeChains Sequence LengthOrganismDetailsImage
DNA-directed RNA polymerase subunit alphaB [auth A],
C [auth B]
347Mycobacterium tuberculosisMutation(s): 0 
Gene Names: rpoA
EC: 2.7.7.6
UniProt
Find proteins for P9WGZ1 (Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv))
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UniProt GroupP9WGZ1
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Entity ID: 3
MoleculeChains Sequence LengthOrganismDetailsImage
DNA-directed RNA polymerase subunit betaD [auth C]1,172Mycobacterium tuberculosisMutation(s): 0 
Gene Names: rpoBMRA_0676
EC: 2.7.7.6
UniProt
Find proteins for P9WGY9 (Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv))
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UniProt GroupP9WGY9
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Entity ID: 4
MoleculeChains Sequence LengthOrganismDetailsImage
DNA-directed RNA polymerase subunit beta'E [auth D]1,318Mycobacterium tuberculosisMutation(s): 0 
Gene Names: rpoCE5M52_05480ERS007665_00591ERS013471_00574ERS023446_00410ERS075361_00813ERS094182_01340F6W99_03342
EC: 2.7.7.6
UniProt
Find proteins for P9WGY7 (Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv))
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UniProt GroupP9WGY7
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Entity ID: 5
MoleculeChains Sequence LengthOrganismDetailsImage
DNA-directed RNA polymerase subunit omegaF [auth E]110Mycobacterium tuberculosisMutation(s): 0 
Gene Names: rpoZERS007703_04032ERS007720_04749ERS094182_01030
EC: 2.7.7.6
UniProt
Find proteins for P9WGY5 (Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv))
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Entity ID: 6
MoleculeChains LengthOrganismImage
DNA (54-MER)G [auth O]54synthetic construct
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Entity ID: 7
MoleculeChains LengthOrganismImage
DNA (54-MER)H [auth P]54synthetic construct
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Entity ID: 8
MoleculeChains LengthOrganismImage
RNA (42-MER)I [auth R]42synthetic construct
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Experimental Data & Validation

Experimental Data

  • Method: ELECTRON MICROSCOPY
  • Resolution: 2.94 Å
  • Aggregation State: PARTICLE 
  • Reconstruction Method: SINGLE PARTICLE 

Structure Validation

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Entry History & Funding Information

Deposition Data


Funding OrganizationLocationGrant Number
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)United States--

Revision History  (Full details and data files)

  • Version 1.0: 2023-03-22
    Type: Initial release
  • Version 1.1: 2023-10-11
    Changes: Data collection, Database references