1887

Abstract

employs two-component systems (TCSs) for survival within its host. The TCS MtrAB is conserved among mycobacteria. The response regulator MtrA is essential in . The genome-wide chromatin immunoprecipitation (ChIP) sequencing performed in this study suggested that MtrA binds upstream of at least 45 genes of , including those involved in cell wall remodelling, stress responses, persistence and regulation of transcription. It binds to the promoter regions and regulates the peptidoglycan hydrolases and , which are required for resuscitation from dormancy. It also regulates the expression of , a critical regulator of the oxidative stress response, and , one-half of the toxin–antitoxin locus . We have identified a new consensus 9 bp loose motif for MtrA binding. Mutational changes in the consensus sequence greatly reduced the binding of MtrA to its newly identified targets. Importantly, we observed that overexpression of a gain-of-function mutant, MtrAY102C, enhanced expression of the aforesaid genes in isolated from macrophages, whereas expression of each of these targets was lower in overexpressing a phosphorylation-defective mutant, MtrAD56N. This result suggests that phosphorylated MtrA (MtrA-P) is required for the expression of its targets in macrophages. Our data have uncovered new MtrA targets that suggest that MtrA is required for a transcriptional response that likely enables to persist within its host and emerge out of dormancy when the conditions are favourable.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000585
2018-01-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/164/1/99.html?itemId=/content/journal/micro/10.1099/mic.0.000585&mimeType=html&fmt=ahah

References

  1. Schnappinger D, Ehrt S, Voskuil MI, Liu Y, Mangan JA et al. Transcriptional adaptation of Mycobacterium tuberculosis within macrophages: insights into the phagosomal environment. J Exp Med 2003; 198:693–704 [View Article][PubMed]
    [Google Scholar]
  2. Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C et al. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 1998; 393:537–544 [View Article][PubMed]
    [Google Scholar]
  3. Bretl DJ, Demetriadou C, Zahrt TC. Adaptation to environmental stimuli within the host: two-component signal transduction systems of Mycobacterium tuberculosis. Microbiol Mol Biol Rev 2011; 75:566–582 [View Article][PubMed]
    [Google Scholar]
  4. Zahrt TC, Deretic V. An essential two-component signal transduction system in Mycobacterium tuberculosis. J Bacteriol 2000; 182:3832–3838 [View Article][PubMed]
    [Google Scholar]
  5. Fol M, Chauhan A, Nair NK, Maloney E, Moomey M et al. Modulation of Mycobacterium tuberculosis proliferation by MtrA, an essential two-component response regulator. Mol Microbiol 2006; 60:643–657 [View Article][PubMed]
    [Google Scholar]
  6. Plocinska R, Purushotham G, Sarva K, Vadrevu IS, Pandeeti EV et al. Septal localization of the Mycobacterium tuberculosis MtrB sensor kinase promotes MtrA regulon expression. J Biol Chem 2012; 287:23887–23899 [View Article][PubMed]
    [Google Scholar]
  7. Rajagopalan M, Dziedzic R, Al Zayer M, Stankowska D, Ouimet MC et al. Mycobacterium tuberculosis origin of replication and the promoter for immunodominant secreted antigen 85B are the targets of MtrA, the essential response regulator. J Biol Chem 2010; 285:15816–15827 [View Article][PubMed]
    [Google Scholar]
  8. Sharma AK, Chatterjee A, Gupta S, Banerjee R, Mandal S et al. MtrA, an essential response regulator of the MtrAB two-component system, regulates the transcription of resuscitation-promoting factor B of Mycobacterium tuberculosis. Microbiology 2015; 161:1271–1281 [View Article][PubMed]
    [Google Scholar]
  9. Winkler ME, Hoch JA. Essentiality, bypass, and targeting of the YycFG (VicRK) two-component regulatory system in Gram-positive bacteria. J Bacteriol 2008; 190:2645–2648 [View Article][PubMed]
    [Google Scholar]
  10. Bisicchia P, Noone D, Lioliou E, Howell A, Quigley S et al. The essential YycFG two-component system controls cell wall metabolism in Bacillus subtilis. Mol Microbiol 2007; 65:180–200 [View Article][PubMed]
    [Google Scholar]
  11. Dubrac S, Bisicchia P, Devine KM, Msadek T. A matter of life and death: cell wall homeostasis and the WalKR (YycGF) essential signal transduction pathway. Mol Microbiol 2008; 70:1307–1322 [View Article][PubMed]
    [Google Scholar]
  12. Kana BD, Mizrahi V. Resuscitation-promoting factors as lytic enzymes for bacterial growth and signaling. FEMS Immunol Med Microbiol 2010; 58:39–50 [View Article][PubMed]
    [Google Scholar]
  13. Downing KJ, Betts JC, Young DI, Mcadam RA, Kelly F et al. Global expression profiling of strains harbouring null mutations reveals that the five rpf-like genes of Mycobacterium tuberculosis show functional redundancy. Tuberculosis 2004; 84:167–179 [View Article][PubMed]
    [Google Scholar]
  14. Russell-Goldman E, Xu J, Wang X, Chan J, Tufariello JM. A Mycobacterium tuberculosis Rpf double-knockout strain exhibits profound defects in reactivation from chronic tuberculosis and innate immunity phenotypes. Infect Immun 2008; 76:4269–4281 [View Article][PubMed]
    [Google Scholar]
  15. Fabret C, Hoch JA. A two-component signal transduction system essential for growth of Bacillus subtilis: implications for anti-infective therapy. J Bacteriol 1998; 180:6375–6383[PubMed]
    [Google Scholar]
  16. Nguyen HT, Wolff KA, Cartabuke RH, Ogwang S, Nguyen L. A lipoprotein modulates activity of the MtrAB two-component system to provide intrinsic multidrug resistance, cytokinetic control and cell wall homeostasis in Mycobacterium. Mol Microbiol 2010; 76:348–364 [View Article][PubMed]
    [Google Scholar]
  17. Valdivia RH, Hromockyj AE, Monack D, Ramakrishnan L, Falkow S. Applications for green fluorescent protein (GFP) in the study of host-pathogen interactions. Gene 1996; 173:47–52 [View Article][PubMed]
    [Google Scholar]
  18. Sanyal S, Banerjee SK, Banerjee R, Mukhopadhyay J, Kundu M. Polyphosphate kinase 1, a central node in the stress response network of Mycobacterium tuberculosis, connects the two-component systems MprAB and SenX3–RegX3 and the extracytoplasmic function sigma factor, sigma E. Microbiology 2013; 159:2074–2086 [View Article][PubMed]
    [Google Scholar]
  19. Rohde KH, Abramovitch RB, Russell DG. Mycobacterium tuberculosis invasion of macrophages: linking bacterial gene expression to environmental cues. Cell Host Microbe 2007; 2:352–364 [View Article][PubMed]
    [Google Scholar]
  20. Langmead B, Trapnell C, Pop M, Salzberg SL. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 2009; 10:R25 [View Article][PubMed]
    [Google Scholar]
  21. Peddireddy V, Doddam SN, Ahmed N. Mycobacterial dormancy systems and host responses in tuberculosis. Front Immunol 2017; 8:84 [View Article][PubMed]
    [Google Scholar]
  22. Satsangi AT, Pandeeti EP, Sarva K, Rajagopalan M, Madiraju MV. Mycobacterium tuberculosis MtrAY102C is a gain-of-function mutant that potentially acts as a constitutively active protein. Tuberculosis 2013; 93:S28–S32 [View Article][PubMed]
    [Google Scholar]
  23. Brocker M, Mack C, Bott M. Target genes, consensus binding site, and role of phosphorylation for the response regulator MtrA of Corynebacterium glutamicum. J Bacteriol 2011; 193:1237–1249 [View Article][PubMed]
    [Google Scholar]
  24. Minch KJ, Rustad TR, Peterson EJ, Winkler J, Reiss DJ et al. The DNA-binding network of Mycobacterium tuberculosis. Nat Commun 2015; 6:5829 [View Article][PubMed]
    [Google Scholar]
  25. Chawla M, Parikh P, Saxena A, Munshi M, Mehta M et al. Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo. Mol Microbiol 2012; 85:1148–1165 [View Article][PubMed]
    [Google Scholar]
  26. Trivedi A, Singh N, Bhat SA, Gupta P, Kumar A. Redox biology of tuberculosis pathogenesis. Adv Microb Physiol 2012; 60:263–324 [View Article][PubMed]
    [Google Scholar]
  27. Rustad TR, Minch KJ, Ma S, Winkler JK, Hobbs S et al. Mapping and manipulating the Mycobacterium tuberculosis transcriptome using a transcription factor overexpression-derived regulatory network. Genome Biol 2014; 15:502 [View Article][PubMed]
    [Google Scholar]
  28. Gupta RK, Srivastava BS, Srivastava R. Comparative expression analysis of rpf-like genes of Mycobacterium tuberculosis H37Rv under different physiological stress and growth conditions. Microbiology 2010; 156:2714–2722 [View Article][PubMed]
    [Google Scholar]
  29. Mukamolova GV, Turapov OA, Young DI, Kaprelyants AS, Kell DB et al. A family of autocrine growth factors in Mycobacterium tuberculosis. Mol Microbiol 2002; 46:623–635 [View Article][PubMed]
    [Google Scholar]
  30. Rickman L, Scott C, Hunt DM, Hutchinson T, Menéndez MC et al. A member of the cAMP receptor protein family of transcription regulators in Mycobacterium tuberculosis is required for virulence in mice and controls transcription of the rpfA gene coding for a resuscitation promoting factor. Mol Microbiol 2005; 56:1274–1286 [View Article][PubMed]
    [Google Scholar]
  31. Raman S, Hazra R, Dascher CC, Husson RN. Transcription regulation by the Mycobacterium tuberculosis alternative sigma factor SigD and its role in virulence. J Bacteriol 2004; 186:6605–6616 [View Article][PubMed]
    [Google Scholar]
  32. Hett EC, Chao MC, Rubin EJ. Interaction and modulation of two antagonistic cell wall enzymes of mycobacteria. PLoS Pathog 2010; 6:e1001020 [View Article][PubMed]
    [Google Scholar]
  33. Rosser A, Stover C, Pareek M, Mukamolova GV. Resuscitation-promoting factors are important determinants of the pathophysiology in Mycobacterium tuberculosis infection. Crit Rev Microbiol 2017; 43:621–630 [View Article][PubMed]
    [Google Scholar]
  34. Pandey DP, Gerdes K. Toxin-antitoxin loci are highly abundant in free-living but lost from host-associated prokaryotes. Nucleic Acids Res 2005; 33:966–976 [View Article][PubMed]
    [Google Scholar]
  35. Korch SB, Contreras H, Clark-Curtiss JE. Three Mycobacterium tuberculosis Rel toxin-antitoxin modules inhibit mycobacterial growth and are expressed in infected human macrophages. J Bacteriol 2009; 191:1618–1630 [View Article][PubMed]
    [Google Scholar]
  36. Koch A, Mizrahi V, Warner DF. The impact of drug resistance on Mycobacterium tuberculosis physiology: what can we learn from rifampicin?. Emerg Microbes Infect 2014; 3:e17 [View Article]
    [Google Scholar]
  37. Li Y, Zeng J, Zhang H, He ZG. The characterization of conserved binding motifs and potential target genes for M. tuberculosis MtrAB reveals a link between the two-component system and the drug resistance of M. smegmatis. BMC Microbiol 2010; 10:242 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000585
Loading
/content/journal/micro/10.1099/mic.0.000585
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error