한빛사논문
- 조회1,868
Abstract
Jee-Hyun Kima,1, Kathryn M. O’Briena,1, Ritu Sharmaa, Helena I. M. Boshoffb, German Rehrena, Sumit Chakrabortyc, Joshua B. Wallacha, Mercedes Monteleonea, Daniel J. Wilsond, Courtney C. Aldrichd, Clifton E. Barry IIIb, Kyu Y. Rheec, Sabine Ehrta, and Dirk Schnappingera,2
aDepartment of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065;
bTuberculosis Research Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892;
cDepartment of Medicine, Weill Cornell Medical College, New York, NY 10065; and
dCenter for Drug Design, University of Minnesota, Minneapolis, MN 55455
Significance
Chronic bacterial infections, such as those caused by Mycobacterium tuberculosis (Mtb), continue to claim the lives of millions of people. New antibiotics are needed to treat these infections, but their development is hindered by a lack of targets whose inhibition quickly eradicates bacterial pathogens and prevents the survival of drug-tolerant persisters. We describe a unique dual-control (DUC) switch that combines repression of transcription and controlled proteolysis to silence gene activities in Mtb. By conditionally inactivating Mtb’s nicotinamide adenine dinucleotide synthetase, we demonstrate that the DUC switch can identify proteins that this pathogen requires for growth and nonreplicating persistence in vitro and during infections. Targeting such proteins holds the promise of yielding drugs that shorten the duration of antibacterial chemotherapies.
Abstract
Antibacterial drug development suffers from a paucity of targets whose inhibition kills replicating and nonreplicating bacteria. The latter include phenotypically dormant cells, known as persisters, which are tolerant to many antibiotics and often contribute to failure in the treatment of chronic infections. This is nowhere more apparent than in tuberculosis caused by Mycobacterium tuberculosis, a pathogen that tolerates many antibiotics once it ceases to replicate. We developed a strategy to identify proteins that Mycobacterium tuberculosis requires to both grow and persist and whose inhibition has the potential to prevent drug tolerance and persister formation. This strategy is based on a tunable dual-control genetic switch that provides a regulatory range spanning three orders of magnitude, quickly depletes proteins in both replicating and nonreplicating mycobacteria, and exhibits increased robustness to phenotypic reversion. Using this switch, we demonstrated that depletion of the nicotinamide adenine dinucleotide synthetase (NadE) rapidly killed Mycobacterium tuberculosis under conditions of standard growth and nonreplicative persistence induced by oxygen and nutrient limitation as well as during the acute and chronic phases of infection in mice. These findings establish the dual-control switch as a robust tool with which to probe the essentiality of Mycobacterium tuberculosis proteins under different conditions, including those that induce antibiotic tolerance, and NadE as a target with the potential to shorten current tuberculosis chemotherapies.
1J.-H.K. and K.M.O. contributed equally to this work.
2To whom correspondence should be addressed.
Author contributions: J.-H.K., K.M.O., R.S., H.I.M.B., G.R., S.C., M.M., C.E.B., K.Y.R., S.E., and D.S. designed research; J.-H.K., K.M.O., R.S., H.I.M.B., G.R., S.C., J.B.W., and M.M. performed research; D.J.W. and C.C.A. contributed new reagents/analytic tools; J.-H.K., K.M.O., R.S., H.I.M.B., G.R., S.C., J.B.W., M.M., C.C.A., C.E.B., K.Y.R., S.E., and D.S. analyzed data; and J.-H.K., K.M.O., C.E.B., K.Y.R., S.E., and D.S. wrote the paper.
논문정보
- Research article
- 2013년 11월 (BRIC 등록일 2013-11-21)
- 국외 연구진
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