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J Biol Chem. 2018 Jun 1;293(22):8379-8393. doi: 10.1074/jbc.RA118.002251. Epub 2018 Apr 9.

The antibiotic cyclomarin blocks arginine-phosphate-induced millisecond dynamics in the N-terminal domain of ClpC1 from Mycobacterium tuberculosis.

Author information

1
From the Institut de Biologie Structurale, University of Grenoble Alpes-CEA, CNRS, IBS, 71 Avenue des Martyrs, CS 10090, 38044 Grenoble Cedex 9, France.
2
the European Molecular Biology Laboratory, 71 avenue des Martyrs, CS 90181, 38042 Grenoble Cedex 9, France.
3
the Institute of Organic Chemistry, Saarland University, Campus C4.2, 66123 Saarbrücken, Germany.
4
the Diseases of the Developing World Discovery Performance Unit, GlaxoSmithKline, Severo Ochoa 2, 28760 Tres Cantos, Madrid, Spain.
5
the Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, and.
6
From the Institut de Biologie Structurale, University of Grenoble Alpes-CEA, CNRS, IBS, 71 Avenue des Martyrs, CS 10090, 38044 Grenoble Cedex 9, France, paul.schanda@ibs.fr.
7
From the Institut de Biologie Structurale, University of Grenoble Alpes-CEA, CNRS, IBS, 71 Avenue des Martyrs, CS 10090, 38044 Grenoble Cedex 9, France, hfraga@med.up.pt.
8
the Departamento de Biomedicina, Faculdade de Medicina and i3S, Instituto de Investigaçào e Inovaçào em Saúde, Universidade do Porto, Alameda Professor Hernàni Monteiro, 4200-319 Porto, Portugal.

Abstract

Mycobacterium tuberculosis can remain dormant in the host, an ability that explains the failure of many current tuberculosis treatments. Recently, the natural products cyclomarin, ecumicin, and lassomycin have been shown to efficiently kill Mycobacterium tuberculosis persisters. Their target is the N-terminal domain of the hexameric AAA+ ATPase ClpC1, which recognizes, unfolds, and translocates protein substrates, such as proteins containing phosphorylated arginine residues, to the ClpP1P2 protease for degradation. Surprisingly, these antibiotics do not inhibit ClpC1 ATPase activity, and how they cause cell death is still unclear. Here, using NMR and small-angle X-ray scattering, we demonstrate that arginine-phosphate binding to the ClpC1 N-terminal domain induces millisecond dynamics. We show that these dynamics are caused by conformational changes and do not result from unfolding or oligomerization of this domain. Cyclomarin binding to this domain specifically blocked these N-terminal dynamics. On the basis of these results, we propose a mechanism of action involving cyclomarin-induced restriction of ClpC1 dynamics, which modulates the chaperone enzymatic activity leading eventually to cell death.

KEYWORDS:

Mycobacterium tuberculosis; antibiotic action; antibiotic resistance; chaperone; natural product; nuclear magnetic resonance (NMR); protease; small-angle X-ray scattering (SAXS)

PMID:
29632076
PMCID:
PMC5986217
DOI:
10.1074/jbc.RA118.002251
[Indexed for MEDLINE]
Free PMC Article

Conflict of interest statement

The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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