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Protein kinases and nitric oxide in the stealthy survival of Mycobacterium tuberculosis

Primary Supervisor: Professor Galina Mukamolova, Department of Respiratory Sciences

Secondary Supervisor: Helen O’Hare, Department of Respiratory Sciences

PhD project title: Protein kinases and nitric oxide in the stealthy survival of Mycobacterium tuberculosis

University of Registration: University of Leicester

Project outline

Tuberculosis remains a major global threat as it affects humans and production animals. Mycobacterium tuberculosis and Mycobacterium bovis, the causative agents of tuberculosis in human and animals, are highly successful pathogens which have sophisticated mechanisms for adaptation to hostile conditions. Nitric oxide (NO) has been long implicated in triggering dormancy in mycobacteria, while serine threonine protein kinases PknB and PknG have been shown to be critical for reactivation from dormancy and regulation of central metabolism. We have recently established that treatment of M. tuberculosis with NO resulted in significant up-regulation of PknB and PknG as well as CwlM and SigH, PknB substrates. Furthermore, we found that over-expression of CwlM stimulated M. tuberculosis survival in activated macrophages. These findings suggest that PknB-mediated phosphorylation is important for mycobacterial survival under nitrosative stress and during macrophage infection. Moreover, PknG is essential for virulence and survival in dormancy.

The proposed project will be focused on investigation of PknB and PknG interplay in NO-treated mycobacteria and during recovery from nitrosative stress. We will also study how depletion of PknB and PknG (as well as their over-expression) impact on mycobacterial survival.

References:

  1. Turapov et al. Two Faces of CwlM, an Essential PknB Substrate, in Mycobacterium tuberculosis. Cell Rep. 2018 Oct 2;25(1):57-67. e5. doi: 10.1016/j.celrep.2018.09.004. https://www.sciencedirect.com/science/article/pii/S2211124718314190?via%3Dihub
  2. Rieck et al. PknG senses amino acid availability to control metabolism and virulence of Mycobacterium tuberculosis. PLoS Path. 13(5): e1006399. https://doi.org/10.1371/journal.ppat.1006399 http://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1006399

BBSRC Strategic Research Priority: Understanding the Rules of Life: Microbiology

Techniques that will be undertaken during the project:

  • Genetic manipulation of mycobacteria
  • generation of deletion and over-expression mutants
  • Growth assays
  • Phosphoproteomic and S-nitrosome analyses
  • SDS-PAGE and immunoblotting
  • Macrophage infection studies

Contact: Professor Galina Mukamolova, University of Leicester