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

Autor:	Delbeau M; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA., Omollo EO; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA., Froom R; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA; Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY 10065, USA., Koh S; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA., Mooney RA; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA., Lilic M; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA., Brewer JJ; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA., Rock J; Laboratory of Host-Pathogen Biology, The Rockefeller University, New York, NY 10065, USA., Darst SA; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA. Electronic address: darst@rockefeller.edu., Campbell EA; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA. Electronic address: campbee@rockefeller.edu., Landick R; Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address: rlandick@wisc.edu.
Jazyk:	angličtina
Zdroj:	Molecular cell [Mol Cell] 2023 May 04; Vol. 83 (9), pp. 1474-1488.e8. Date of Electronic Publication: 2023 Apr 27.
DOI:	10.1016/j.molcel.2023.04.007
Abstrakt:	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. Competing Interests: Declaration of interests R.L. is a member of the Molecular Cell advisory board. (Copyright © 2023 Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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