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TitleStructural and functional basis of the universal transcription factor NusG pro-pausing activity in Mycobacterium tuberculosis.
Journal, issue, pagesMol Cell, Vol. 83, Issue 9, Page 1474-11488.e8, Year 2023
Publish dateMay 4, 2023
AuthorsMadeleine Delbeau / Expery O Omollo / Ruby Froom / Steven Koh / Rachel A Mooney / Mirjana Lilic / Joshua J Brewer / Jeremy Rock / Seth A Darst / Elizabeth A Campbell / Robert Landick /
PubMed AbstractTranscriptional 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 ...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.
External linksMol Cell / PubMed:37116494 / PubMed Central
MethodsEM (single particle)
Resolution2.9 - 3.71 Å
Structure data

EMDB-27935, PDB-8e74:
Mycobacterium tuberculosis RNAP paused elongation complex with NusG transcription factor
Method: EM (single particle) / Resolution: 2.94 Å

EMDB-27938, PDB-8e79:
Mycobacterium tuberculosis RNAP paused elongation complex with Escherichia coli NusG transcription factor
Method: EM (single particle) / Resolution: 3.71 Å

EMDB-27942, PDB-8e82:
Mycobacterium tuberculosis RNAP elongation complex with NusG transcription factor
Method: EM (single particle) / Resolution: 3.03 Å

EMDB-27944, PDB-8e8m:
Mycobacterium tuberculosis RNAP paused elongation complex
Method: EM (single particle) / Resolution: 3.13 Å

EMDB-27956, PDB-8e95:
Mycobacterium tuberculosis RNAP elongation complex
Method: EM (single particle) / Resolution: 2.9 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

Source
  • mycobacterium tuberculosis (bacteria)
  • synthetic construct (others)
  • escherichia coli (E. coli)
KeywordsTRANSCRIPTION/DNA/RNA / RNA polymerase / transcription factor / elongation / pausing / transcription-transferase-DNA complex / TRANSCRIPTION-DNA-RNA complex / TRANSCRIPTION / TRANSFERASE/DNA/RNA / TRANSFERASE-DNA-RNA complex

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