Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis

Virginie Molle, Gulcin Gulten, Catherine Vilchèze, Romain Veyron-Churlet, Isabelle Zanella-Cléon, James C. Sacchettini, William R. Jacobs, Laurent Kremer

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The remarkable survival ability of Mycobacterium tuberculosis in infected hosts is related to the presence of cell wall-associated mycolic acids. Despite their importance, the mechanisms that modulate expression of these lipids in response to environmental changes are unknown. Here we demonstrate that the enoyl-ACP reductase activity of InhA, an essential enzyme of the mycolic acid biosynthetic pathway and the primary target of the anti-tubercular drug isoniazid, is controlled via phosphorylation. Thr-266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr-266 phosphorylation was demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity was severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Importantly, introduction of inhA_T266D/E failed to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. This study suggests that phosphorylation of InhA may represent an unusual mechanism that allows M. tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development.

Original languageEnglish (US)
Pages (from-to)1591-1605
Number of pages15
JournalMolecular Microbiology
Volume78
Issue number6
DOIs
StatePublished - Dec 2010

Fingerprint

Mycolic Acids
Mycobacterium tuberculosis
Phosphorylation
Isoniazid
Growth
Oxidoreductases
Mycobacterium smegmatis
Biosynthetic Pathways
Pharmaceutical Preparations
NAD
Cell Wall
Tuberculosis
Phosphotransferases
Lipids
Enzymes
In Vitro Techniques

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Molle, V., Gulten, G., Vilchèze, C., Veyron-Churlet, R., Zanella-Cléon, I., Sacchettini, J. C., ... Kremer, L. (2010). Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis. Molecular Microbiology, 78(6), 1591-1605. https://doi.org/10.1111/j.1365-2958.2010.07446.x

Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis. / Molle, Virginie; Gulten, Gulcin; Vilchèze, Catherine; Veyron-Churlet, Romain; Zanella-Cléon, Isabelle; Sacchettini, James C.; Jacobs, William R.; Kremer, Laurent.

In: Molecular Microbiology, Vol. 78, No. 6, 12.2010, p. 1591-1605.

Research output: Contribution to journalArticle

Molle, V, Gulten, G, Vilchèze, C, Veyron-Churlet, R, Zanella-Cléon, I, Sacchettini, JC, Jacobs, WR & Kremer, L 2010, 'Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis', Molecular Microbiology, vol. 78, no. 6, pp. 1591-1605. https://doi.org/10.1111/j.1365-2958.2010.07446.x
Molle, Virginie ; Gulten, Gulcin ; Vilchèze, Catherine ; Veyron-Churlet, Romain ; Zanella-Cléon, Isabelle ; Sacchettini, James C. ; Jacobs, William R. ; Kremer, Laurent. / Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis. In: Molecular Microbiology. 2010 ; Vol. 78, No. 6. pp. 1591-1605.
@article{60dd04698e614909a6c35e7a7d49df0a,
title = "Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis",
abstract = "The remarkable survival ability of Mycobacterium tuberculosis in infected hosts is related to the presence of cell wall-associated mycolic acids. Despite their importance, the mechanisms that modulate expression of these lipids in response to environmental changes are unknown. Here we demonstrate that the enoyl-ACP reductase activity of InhA, an essential enzyme of the mycolic acid biosynthetic pathway and the primary target of the anti-tubercular drug isoniazid, is controlled via phosphorylation. Thr-266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr-266 phosphorylation was demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity was severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Importantly, introduction of inhA_T266D/E failed to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. This study suggests that phosphorylation of InhA may represent an unusual mechanism that allows M. tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development.",
author = "Virginie Molle and Gulcin Gulten and Catherine Vilch{\`e}ze and Romain Veyron-Churlet and Isabelle Zanella-Cl{\'e}on and Sacchettini, {James C.} and Jacobs, {William R.} and Laurent Kremer",
year = "2010",
month = "12",
doi = "10.1111/j.1365-2958.2010.07446.x",
language = "English (US)",
volume = "78",
pages = "1591--1605",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis

AU - Molle, Virginie

AU - Gulten, Gulcin

AU - Vilchèze, Catherine

AU - Veyron-Churlet, Romain

AU - Zanella-Cléon, Isabelle

AU - Sacchettini, James C.

AU - Jacobs, William R.

AU - Kremer, Laurent

PY - 2010/12

Y1 - 2010/12

N2 - The remarkable survival ability of Mycobacterium tuberculosis in infected hosts is related to the presence of cell wall-associated mycolic acids. Despite their importance, the mechanisms that modulate expression of these lipids in response to environmental changes are unknown. Here we demonstrate that the enoyl-ACP reductase activity of InhA, an essential enzyme of the mycolic acid biosynthetic pathway and the primary target of the anti-tubercular drug isoniazid, is controlled via phosphorylation. Thr-266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr-266 phosphorylation was demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity was severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Importantly, introduction of inhA_T266D/E failed to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. This study suggests that phosphorylation of InhA may represent an unusual mechanism that allows M. tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development.

AB - The remarkable survival ability of Mycobacterium tuberculosis in infected hosts is related to the presence of cell wall-associated mycolic acids. Despite their importance, the mechanisms that modulate expression of these lipids in response to environmental changes are unknown. Here we demonstrate that the enoyl-ACP reductase activity of InhA, an essential enzyme of the mycolic acid biosynthetic pathway and the primary target of the anti-tubercular drug isoniazid, is controlled via phosphorylation. Thr-266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr-266 phosphorylation was demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity was severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Importantly, introduction of inhA_T266D/E failed to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. This study suggests that phosphorylation of InhA may represent an unusual mechanism that allows M. tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development.

UR - http://www.scopus.com/inward/record.url?scp=78650031727&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78650031727&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2958.2010.07446.x

DO - 10.1111/j.1365-2958.2010.07446.x

M3 - Article

C2 - 21143326

AN - SCOPUS:78650031727

VL - 78

SP - 1591

EP - 1605

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 6

ER -