Characterization of a two-component system, devR-devS, of Mycobacterium tuberculosis

2000 ◽  
Vol 80 (3) ◽  
pp. 141-159 ◽  
Author(s):  
N. Dasgupta ◽  
V. Kapur ◽  
K.K. Singh ◽  
T.K. Das ◽  
S. Sachdeva ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Two Component System ◽  
Component System ◽  
Two Component

scholarly journals Kinetic Characterization of the WalRKSpn (VicRK) Two-Component System of Streptococcus pneumoniae: Dependence of WalKSpn (VicK) Phosphatase Activity on Its PAS Domain

2010 ◽  
Vol 192 (9) ◽  
pp. 2346-2358 ◽  
Author(s):  
Alina D. Gutu ◽  
Kyle J. Wayne ◽  
Lok-To Sham ◽  
Malcolm E. Winkler
Keyword(s):  
Amino Acids ◽  
Streptococcus Pneumoniae ◽  
Phosphatase Activity ◽  
Pas Domain ◽  
Kinetic Characterization ◽  
Two Component System ◽  
Histidine Kinases ◽  
Component System ◽  
Two Component

ABSTRACT The WalRK two-component system plays important roles in maintaining cell wall homeostasis and responding to antibiotic stress in low-GC Gram-positive bacteria. In the major human pathogen, Streptococcus pneumoniae, phosphorylated WalR Spn (VicR) response regulator positively controls the transcription of genes encoding the essential PcsB division protein and surface virulence factors. WalR Spn is phosphorylated by the WalK Spn (VicK) histidine kinase. Little is known about the signals sensed by WalK histidine kinases. To gain information about WalK Spn signal transduction, we performed a kinetic characterization of the WalRK Spn autophosphorylation, phosphoryltransferase, and phosphatase reactions. We were unable to purify soluble full-length WalK Spn . Consequently, these analyses were performed using two truncated versions of WalK Spn lacking its single transmembrane domain. The longer version (Δ35 amino acids) contained most of the HAMP domain and the PAS, DHp, and CA domains, whereas the shorter version (Δ195 amino acids) contained only the DHp and CA domains. The autophosphorylation kinetic parameters of Δ35 and Δ195 WalK Spn were similar [Km (ATP) ≈ 37 μM; k cat ≈ 0.10 min−1] and typical of those of other histidine kinases. The catalytic efficiency of the two versions of WalK Spn ∼P were also similar in the phosphoryltransfer reaction to full-length WalR Spn . In contrast, absence of the HAMP-PAS domains significantly diminished the phosphatase activity of WalK Spn for WalR Spn ∼P. Deletion and point mutations confirmed that optimal WalK Spn phosphatase activity depended on the PAS domain as well as residues in the DHp domain. In addition, these WalK Spn DHp domain and ΔPAS mutations led to attenuation of virulence in a murine pneumonia model.

scholarly journals Mycobacterium tuberculosis PhoY Proteins Promote Persister Formation by Mediating Pst/SenX3-RegX3 Phosphate Sensing

mBio ◽  
2017 ◽  
Vol 8 (4) ◽  
Author(s):  
Sarah B. Namugenyi ◽  
Alisha M. Aagesen ◽  
Sarah R. Elliott ◽  
Anna D. Tischler
Keyword(s):  
Signal Transduction ◽  
Mycobacterium Tuberculosis ◽  
System Component ◽  
Signal Transduction System ◽  
Two Component System ◽  
Component System ◽  
Phosphate Sensing ◽  
Two Component ◽  
Pst System

ABSTRACT The Mycobacterium tuberculosis phosphate-specific transport (Pst) system controls gene expression in response to phosphate availability by inhibiting the activation of the SenX3-RegX3 two-component system under phosphate-rich conditions, but the mechanism of communication between these systems is unknown. In Escherichia coli, inhibition of the two-component system PhoR-PhoB under phosphate-rich conditions requires both the Pst system and PhoU, a putative adaptor protein. E. coli PhoU is also involved in the formation of persisters, a subpopulation of phenotypically antibiotic-tolerant bacteria. M. tuberculosis encodes two PhoU orthologs, PhoY1 and PhoY2. We generated phoY single- and double-deletion mutants and examined the expression of RegX3-regulated genes by quantitative reverse transcription-PCR (qRT-PCR). Gene expression was increased only in the ΔphoY1 ΔphoY2 double mutant and could be restored to the wild-type level by complementation with either phoY1 or phoY2 or by deletion of regX3. These data suggest that the PhoY proteins function redundantly to inhibit SenX3-RegX3 activation. We analyzed the frequencies of antibiotic-tolerant persister variants in the phoY mutants using several antibiotic combinations. Persister frequency was decreased at least 40-fold in the ΔphoY1 ΔphoY2 mutant compared to the frequency in the wild type, and this phenotype was RegX3 dependent. A ΔpstA1 mutant lacking a Pst system transmembrane component exhibited a similar RegX3-dependent decrease in persister frequency. In aerosol-infected mice, the ΔphoY1 ΔphoY2 and ΔpstA1 mutants were more susceptible to treatment with rifampin but not isoniazid. Our data demonstrate that disrupting phosphate sensing mediated by the PhoY proteins and the Pst system enhances the susceptibility of M. tuberculosis to antibiotics both in vitro and during infection. IMPORTANCE Persister variants, subpopulations of bacteria that are phenotypically antibiotic tolerant, contribute to the lengthy treatment times required to cure Mycobacterium tuberculosis infection, but the molecular mechanisms governing their formation and maintenance are poorly characterized. Here, we demonstrate that a phosphate-sensing signal transduction system, comprising the Pst phosphate transporter, the two-component system SenX3-RegX3, and functionally redundant PhoY proteins that mediate signaling between Pst and SenX3-RegX3, influences persister formation. Activation of RegX3 by deletion of the phoY genes or a Pst system component resulted in decreased persister formation in vitro. Activated RegX3 also limited persister formation during growth under phosphate-limiting conditions. Importantly, increased susceptibility to the front-line drug rifampin was also observed in a mouse infection model. Thus, the M. tuberculosis phosphate-sensing signal transduction system contributes to antibiotic tolerance and is a potential target for the development of novel therapeutics that may shorten the duration of tuberculosis treatment. IMPORTANCE Persister variants, subpopulations of bacteria that are phenotypically antibiotic tolerant, contribute to the lengthy treatment times required to cure Mycobacterium tuberculosis infection, but the molecular mechanisms governing their formation and maintenance are poorly characterized. Here, we demonstrate that a phosphate-sensing signal transduction system, comprising the Pst phosphate transporter, the two-component system SenX3-RegX3, and functionally redundant PhoY proteins that mediate signaling between Pst and SenX3-RegX3, influences persister formation. Activation of RegX3 by deletion of the phoY genes or a Pst system component resulted in decreased persister formation in vitro. Activated RegX3 also limited persister formation during growth under phosphate-limiting conditions. Importantly, increased susceptibility to the front-line drug rifampin was also observed in a mouse infection model. Thus, the M. tuberculosis phosphate-sensing signal transduction system contributes to antibiotic tolerance and is a potential target for the development of novel therapeutics that may shorten the duration of tuberculosis treatment.

scholarly journals The prrAB Two-Component System Is Essential for Mycobacterium tuberculosis Viability and Is Induced under Nitrogen-Limiting Conditions

2011 ◽  
Vol 194 (2) ◽  
pp. 354-361 ◽  
Author(s):  
S. E. Haydel ◽  
V. Malhotra ◽  
G. L. Cornelison ◽  
J. E. Clark-Curtiss
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Two Component System ◽  
Component System ◽  
Two Component

Characterization of a Novel Two-Component System in Burkholderia cenocepacia

2014 ◽  
Vol 70 (4) ◽  
pp. 556-561 ◽  
Author(s):  
Callie R. Merry ◽  
Michael Perkins ◽  
Lin Mu ◽  
Bridget K. Peterson ◽  
Rebecca W. Knackstedt ◽  
...  
Keyword(s):  
Burkholderia Cenocepacia ◽  
Two Component System ◽  
Component System ◽  
Two Component

scholarly journals The MprB Extracytoplasmic Domain Negatively Regulates Activation of the Mycobacterium tuberculosis MprAB Two-Component System

2013 ◽  
Vol 196 (2) ◽  
pp. 391-406 ◽  
Author(s):  
D. J. Bretl ◽  
T. M. Bigley ◽  
S. S. Terhune ◽  
T. C. Zahrt
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Two Component System ◽  
Component System ◽  
Two Component

scholarly journals Corrigendum: MtrA, an essential response regulator of the MtrAB two-component system, regulates the transcription of resuscitation-promoting factor B of Mycobacterium tuberculosis

Microbiology ◽  
2020 ◽  
Vol 166 (7) ◽  
pp. 679-679
Author(s):  
Arun Kumar Sharma ◽  
Ayan Chatterjee ◽  
Shamba Gupta ◽  
Rajdeep Banerjee ◽  
Sukhendu Mandal ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Response Regulator ◽  
Two Component System ◽  
Factor B ◽  
Component System ◽  
Two Component

The Mycobacterium tuberculosis PhoPR two-component system regulates genes essential for virulence and complex lipid biosynthesis

2006 ◽  
Vol 60 (2) ◽  
pp. 312-330 ◽  
Author(s):  
Shaun B. Walters ◽  
Eugenie Dubnau ◽  
Irina Kolesnikova ◽  
Francoise Laval ◽  
Mamadou Daffe ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Lipid Biosynthesis ◽  
Two Component System ◽  
Component System ◽  
Two Component ◽  
Complex Lipid

Interaction analysis of TcrX/Y two component system from Mycobacterium tuberculosis

Biochimie ◽  
2010 ◽  
Vol 92 (3) ◽  
pp. 263-272 ◽  
Author(s):  
Monolekha Bhattacharya ◽  
Ashis Biswas ◽  
Amit Kumar Das
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Interaction Analysis ◽  
Two Component System ◽  
Component System ◽  
Two Component

Genome-wide characterization of two-component system (TCS) genes in melon (Cucumis melo L.)

2020 ◽  
Vol 151 ◽  
pp. 197-213 ◽  
Author(s):  
Panjing Liu ◽  
Shuoshuo Wang ◽  
Xiangfei Wang ◽  
Xiaoyu Yang ◽  
Qiang Li ◽  
...  
Keyword(s):  
Cucumis Melo ◽  
Two Component System ◽  
Component System ◽  
Genome Wide ◽  
Two Component ◽  
Cucumis Melo L
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