CoChat
Sign Up
Cited by 13Tuberculosis Research and EpidemiologyBacterial Genetics and BiotechnologyMycobacterium research and diagnosis

A Serine/threonine kinase PknL , is involved in the adaptive response of Mycobacterium tuberculosis

Open access
Refaya, Ahmed Kabir, Sharma, Divakar, Kumar, Virendra, Bisht, Deepa, Narayanan, Sujatha·2016·Microbiological Research
DOI 10.1016/j.micres.2016.02.005
Open this paper in CoChat — free account

Abstract

Mycobacterium tuberculosis adapts itself to various environmental stress conditions to thrive inside the phagosome for establishing a chronic infection. Serine/threonine protein kinases (STPKs) play a major role in the physiology and pathogenesis of Mycobacterium tuberculosis. Some of these STPKs are involved in regulating the growth of the mycobacterium under nutrient stress and starvation conditions. In this study, we have investigated the role of PknL, a STPK in the adaptive responses of M. tuberculosis by conditional inactivation of the gene using antisense technology. The inhibition of PknL in the knockdown strain was validated by RT-PCR. The in vitro growth kinetics of M. tuberculosis strain following inhibition of PknL was found to be bacteriostatic. The knock down strain of PknL exhibited a better survival in pH 5.5 when compared to its growth in pH 7.0. Similarly, it also exhibited more resistance to both SDS(0.01%) and Lysozyme stress (2.5mg/ml), indicating that loss of PknL enhances the growth of mycobacterium under stress conditions. SEM pictographs also represent an increase in the cell length of the knock down strain compared to Wild type stressing its role in cellular integrity. Lastly, the proteome analysis of differentially expressing PknL strains by 2D gel electrophoresis and mass spectrometry identified 19 differentially expressed proteins. Our findings have shown that PknL plays an important role in sensing the host environment and adapting itself in slowing down the growth of the pathogen and persisting within the host.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.micres.2016.02.005

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • As these STPKs are associated with stasis conditions, their phosphorylation of FAS-II enzymes reduces mycolic acid synthesis during stasis and chronic phases of infection.

    “As these STPKs are associated with stasis conditions14,72,94,95, we predict that their phosphorylation of FAS-​II enzymes reduces mycolic acid synthesis during stasis and chronic phases of infection.”

    From The mycobacterial cell envelope — a moving target (2020) · cites this paper as [95]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 40

  1. Bisht, D., Singhal, N., Sharma, P., Venkatesan, K., 2007. An improved sample preparation method for analyzing mycobacterial proteins in two-dimensional gels. Biochem. Biokhimiia 72 (6), 672-674.
  2. Canova, M.J., Veyron-Churlet, R., Zanella-Cleon, I., Cohen-Gonsaud, M., Cozzone, A.J., Becchi, M., Kremer, L., Molle, V., 2008. The Mycobacterium tuberculosis serine/threonine kinase PknL phosphorylates Rv2175c: mass spectrometric profiling of the activation loop phosphorylation sites and their role in the recruitment of Rv2175c. Proteomics 8 (3), 521-533.
  3. Carroll, P., Faray-Kele, M.C., Parish, T., 2011. Identifying vulnerable pathways in Mycobacterium tuberculosis by using a knockdown approach. Appl. Environ. Microbiol. 77 (14), 5040-5043.
  4. Cole, S.T., Brosch, R., Parkhill, J., Garnier, T., Churcher, C., Harris, D., Gordon, S.V., Eiglmeier, K., Gas, S., Barry 3rd, C.E., Tekaia, F., Badcock, K., Basham, D., Brown, D., Chillingworth, T., Connor, R., Davies, R., Devlin, K., Feltwell, T., Gentles, S., Hamlin, N., Holroyd, S., Hornsby, T., Jagels, K., Krogh, A., McLean, J., Moule, S., Murphy, L., Oliver, K., Osborne, J., Quail, M.A., Rajandream, M.A., Rogers, J., Rutter, S., Seeger, K., Skelton, J., Squares, R., Squares, S., Sulston, J.E., Taylor, K., Whitehead, S., Barrell, B.G., 1998. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393 (6685), 537-544.
  5. Cowley, S., Ko, M., Pick, N., Chow, R., Downing, K.J., Gordhan, B.G., Betts, J.C., Mizrahi, V., Smith, D.A., Stokes, R.W., Av-Gay, Y., 2004. The Mycobacterium tuberculosis protein serine/threonine kinase PknG is linked to cellular glutamate/glutamine levels and is important for growth in vivo. Mol. Microbiol. 52 (6), 1691-1702.
  6. Dahl, J.L., Kraus, C.N., Boshoff, H.I., Doan, B., Foley, K., Avarbock, D., Kaplan, G., Mizrahi, V., Rubin, H., Barry, C.E., 2003. The role of RelMtb-mediated adaptation to stationary phase in long-term persistence of Mycobacterium tuberculosis in mice. Proc. Natl. Acad. Sci. U. S. A. 100 (17), 10026-10031.
  7. Deol, P., Vohra, R., Saini, A.K., Singh, A., Chandra, H., Chopra, P., Das, T.K., Tyagi, A.K., Singh, Y., 2005. Role of Mycobacterium tuberculosis Ser/Thr kinase PknF: implications in glucose transport and cell division. J. Bacteriol. 187 (10), 3415-3420.
  8. Deutscher, J., Saier Jr., M.H., 2005. Ser/Thr/Tyr protein phosphorylation in bacteria-for long time neglected, now well established. J. Mol. Microbiol. Biotechnol. 9 (3-4), 125-131.
  9. Fernandez, P., Saint-Joanis, B., Barilone, N., Jackson, M., Gicquel, B., Cole, S.T., Alzari, P.M., 2006. The Ser/Thr protein kinase PknB is essential for sustaining mycobacterial growth. J. Bacteriol. 188 (22), 7778-7784.
  10. Geiman, D.E., Raghunand, T.R., Agarwal, N., Bishai, W.R., 2006. Differential gene expression in response to exposure to antimycobacterial agents and other stress conditions among seven Mycobacterium tuberculosi s whiB-like genes. Antimicrob. Agents Chemother. 50 (8), 2836-2841.
  11. Gopalaswamy, R., Narayanan, S., Chen, B., Jacobs, W.R., Av-Gay, Y., 2009. The serine/threonine protein kinase PknI controls the growth of Mycobacterium tuberculosis upon infection. FEMS Microbiol. Lett. 295 (1), 23-29.
  12. Greenstein, A.E., Grundner, C., Echols, N., Gay, L.M., Lombana, T.N., Miecskowski, C.A., Pullen, K.E., Sung, P.Y., Alber, T., 2005. Structure/function studies of Ser/Thr and Tyr protein phosphorylation in Mycobacterium tuberculosis . J. Mol. Microbiol. Biotechnol. 9 (3-4), 167-181.
  13. Griffin, J.E., Gawronski, J.D., Dejesus, M.A., Ioerger, T.R., Akerley, B.J., Sassetti, C.M., 2011. High-resolution phenotypic profiling defines genes essential for mycobacterial growth and cholesterol catabolism. PLoS Pathog. 7 (9), e1002251.
  14. J., Bargatze, R.F., Harmsen, A.G., Crouch, E.C., Schlesinger, L.S., 2006. Mycobacterium tuberculosis binding to human surfactant proteins A and D, fibronectin, and small airway epithelial cells under shear conditions. Infect. Immun. 74 (6), 3587-3596.
  15. Harth, G., Maslesa-Galic, S., Tullius, M.V., Horwitz, M.A., 2005. All four Mycobacterium tuberculosis glnA genes encode glutamine synthetase activities but only GlnA1 is abundantly expressed and essential for bacterial homeostasis. Mol. Microbiol. 58 (4), 1157-1172.
  16. Jagannathan, V., Kaur, P., Datta, S., 2010. Polyphosphate kinase from M. tuberculosis : an interconnect between the genetic and biochemical role. PLoS One 5 (12), e14336.
  17. Jayakumar, D., Jacobs Jr., W.R., Narayanan, S., 2008. Protein kinase E of Mycobacterium tuberculosis has a role in the nitric oxide stress response and apoptosis in a human macrophage model of infection. Cell Microbiol. 10 (2), 365-374.
  18. Kang, J.L., Lee, H.W., Kim, H.J., Lee, H.S., Castranova, V., Lim, C.M., Koh, Y., 2005. Inhibition of SRC tyrosine kinases suppresses activation of nuclear factor-kappaB, and serine and tyrosine phosphorylation of IkappaB-alpha in lipopolysaccharide-stimulated raw 264.7 macrophages. J. Toxicol. Environ. Health A 68 (19), 1643-1662.
  19. Kaur, P., Agarwal, S., Datta, S., 2009. Delineating bacteriostatic and bactericidal targets in mycobacteria using IPTG inducible antisense expression. PLoS One 4 (6), e5923.
  20. Lamichhane, G., Zignol, M., Blades, N.J., Geiman, D.E., Dougherty, A., Grosset, J., Broman, K.W., Bishai, W.R., 2003. A postgenomic method for predicting essential genes at subsaturation levels of mutagenesis: application to Mycobacterium tuberculosis . Proc. Natl. Acad. Sci. U. S. A. 100 (12), 7213-7218.
  21. Leistikow, R.L., Morton, R.A., Bartek, I.L., Frimpong, I., Wagner, K., Voskuil, M.I., 2010. The Mycobacterium tuberculosis DosR regulon assists in metabolic homeostasis and enables rapid recovery from nonrespiring dormancy. J. Bacteriol. 192 (6), 1662-1670.
  22. LH, M., 2001. Cell wall deficient forms. In: Stealth Pathogens 3rd edition., pp. 189-197. http://refhub.elsevier.com/S0944-5013(1630010-6/sbref0150)
  23. Livak, K.J., Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25 (4), 402-408.
  24. Malhotra, V., Arteaga-Cortes, L.T., Clay, G., Clark-Curtiss, J.E., 2010. Mycobacterium tuberculosis protein kinase K confers survival advantage during early infection in mice and regulates growth in culture and during persistent infection: implications for immune modulation. Microbiology 156 (Pt. 9), 2829-2841. http://refhub.elsevier.com/S0944-5013(1630010-6/sbref0160)
  25. Molle, V., Kremer, L., 2010. Division and cell envelope regulation by Ser/Thr phosphorylation: Mycobacterium shows the way. Mol. Microbiol. 75 (5), 1064-1077.
  26. Mwandumba, H.C., Russell, D.G., Nyirenda, M.H., Anderson, J., White, S.A., et al., 2004. Mycobacterium tuberculosis resides in nonacidified vacuoles in endocytically competent alveolar macrophages from patients with tuberculosis and HIV infection. J. Immunol. 172 (7), 4592-4598.
  27. Narayan, A., Sachdeva, P., Sharma, K., Saini, A.K., Tyagi, A.K., Singh, Y., 2007. Serine threonine protein kinases of mycobacterial genus: phylogeny to function. Physiol. Genom. 29 (1), 66-75.
  28. Ortega, C., Liao, R., Anderson, L.N., Rustad, T., Ollodart, A.R., Wright, A.T., Sherman, D.R., Grundner, C., 2014. Mycobacterium tuberculosis Ser/Thr protein kinase B mediates an oxygen-dependent replication switch. PLoS Biol. 12 (1), e1001746.
  29. Papavinasasundaram, K.G., Chan, B., Chung, J.H., Colston, M.J., Davis, E.O., Av-Gay, Y., 2005. Deletion of the Mycobacterium tuberculosi s pknH gene confers a higher bacillary load during the chronic phase of infection in BALB/c mice. J. Bacteriol. 187 (16), 5751-5760.
  30. Parida, B.K., Douglas, T., Nino, C., Dhandayuthapani, S., 2005. Interactions of anti-sigma factor antagonists of Mycobacterium tuberculosis in the yeast two-hybrid system. Tuberculosis (Edinb) 85 (5-6), 347-355.
  31. Pethe, K.S.D., Alonso, S., Anderson, J., Wang, C., et al., 2004. Isolation of Mycobacterium tuberculosis mutants defective in the arrest of phagosome maturation. Proc. Natl. Acad. Sci. U. S. A. 101 (37), 13642-13647.
  32. Primm, T.P., Andersen, S.J., Mizrahi, V., Avarbock, D., Rubin, H., Barry, C.E., 2000. The stringent response of Mycobacterium tuberculosis is required for long-term survival. J. Bacteriol. 182 (17), 4889-4898.
  33. Prisic, S., Dankwa, S., Schwartz, D., Chou, M.F., Locasale, J.W., Kang, C.M., Bemis, G., Church, G.M., Steen, H., Husson, R.N., 2010. Extensive phosphorylation with overlapping specificity by Mycobacterium tuberculosis serine/threonine protein kinases. Proc. Natl. Acad. Sci. U. S. A. 107 (16), 7521-7526.
  34. Rohde, K.H., Abramovitch, R.B., Russell, D.G., 2007. Mycobacterium tuberculosis invasion of macrophages: linking bacterial gene expression to environmental cues. Cell Host Microb. 2 (5), 352-364.
  35. Sassetti, C.M., Boyd, D.H., Rubin, E.J., 2003. Genes required for mycobacterial growth defined by high density mutagenesis. Mol. Microbiol. 48 (1), 77-84. http://refhub.elsevier.com/S0944-5013(1630010-6/sbref0215)
  36. Scherr, N., Muller, P., Perisa, D., Combaluzier, B., Jeno, P., Pieters, J., 2009. Survival of pathogenic mycobacteria in macrophages is mediated through autophosphorylation of protein kinase G. J. Bacteriol. 191 (14), 4546-4554.
  37. Sharma, D., Kumar, B., Lata, M., Joshi, B., Venkatesan, K., Shukla, S., Bisht, D., 2015. Comparative proteomic analysis of aminoglycosides resistant and susceptible mycobacterium tuberculosis clinical isolates for exploring potential drug targets. PLoS One 10 (10), e0139414.
  38. Singhal, N., Sharma, P., Kumar, M., Joshi, B., Bisht, D., 2012. Analysis of intracellular expressed proteins of Mycobacterium tuberculosis clinical isolates. Proteome Sci. 10 (1), 14.
  39. Tan, S., Sukumar, N., Abramovitch, R.B., Parish, T., Russell, D.G., 2013. Mycobacterium tuberculosis responds to chloride and pH as synergistic cues to the immune status of its host cell. PLoS Pathog. 9 (4), e1003282.
  40. Walters, S.B., Dubnau, E., Kolesnikova, I., Laval, F., Daffe, M., Smith, I., 2006. The Mycobacterium tuberculosis PhoPR two-component system regulates genes essential for virulence and complex lipid biosynthesis. Mol. Microbiol. 60 (2), 312-330.

Cited by

Sorted by citations

Authors
Refaya, A. K., Sharma, D., Kumar, V., Bisht, D., & Narayanan, S.
Affiliations
  • National Institute of Research in Tuberculosis (IN)
  • National JALMA Institute for Leprosy & Other Mycobacterial Diseases (IN)
Corresponding author
Narayanan, Sujatha
ORCID
Venue
Microbiological Research
Publisher
Elsevier BV
Published
2016-09
Volume
190
Pages
1-11
ISSN
0944-5013 (ISSN-L)
DOI
10.1016/j.micres.2016.02.005
OpenAlex
W2346594768
Cited by
13
References
2
Citations by year
2026: 1 · 2024: 2 · 2021: 3 · 2019: 2 · 2018: 4 · 2016: 1
FWCI
2.16
Type
journal article
Language
en
Open access
Licence
Locations
  • Microbiological Research (publishedVersion)
  • PubMed (publishedVersion)
  • NIRT Institutional Scholarship Repository (National Institute of Research in Tuberculosis) (acceptedVersion)
Funders
Indian Council of Medical Research
Topics
  • Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
Keywords
serine/threonine kinase, Mycobacterium tuberculosis, adaptive response, phagosome, acid stress, lysozyme resistance, SDS resistance, bacterial growth kinetics, cell morphology, differentially expressed proteins, conditional gene knockdown, host-pathogen interaction
MeSH terms
Hydrogen-Ion Concentration, Microscopy, Electron, Scanning, Muramidase, Mycobacterium tuberculosis, Sodium Dodecyl Sulfate, Mass Spectrometry, Stress, Physiological, Electrophoresis, Gel, Two-Dimensional, Protein Serine-Threonine Kinases, Proteome, Gene Expression Profiling, RNA Interference, Microbial Viability, Gene Knockdown Techniques

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review