scholarly journals Allosteric mechanism of signal transduction in the two-component system histidine kinase PhoQ

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Bruk Mensa ◽  
Nicholas F Polizzi ◽  
Kathleen S Molnar ◽  
Andrew M Natale ◽  
Thomas Lemmin ◽  
...  
Keyword(s):  
Histidine Kinase ◽  
Equilibrium Constants ◽  
Kinase Domain ◽  
Domain Model ◽  
Bistable System ◽  
E Coli ◽  
Allosteric Communication ◽  
Two Component ◽  
Cytoplasmic Kinase Domain ◽  
Semi Empirical

Transmembrane signaling proteins couple extracytosolic sensors to cytosolic effectors. Here, we examine how binding of Mg2+ to the sensor domain of an E. coli two component histidine kinase (HK), PhoQ, modulates its cytoplasmic kinase domain. We use cysteine-crosslinking and reporter-gene assays to simultaneously and independently probe the signaling state of PhoQ's sensor and autokinase domains in a set of over 30 mutants. Strikingly, conservative single-site mutations distant from the sensor or catalytic site strongly influence PhoQ's ligand-sensitivity as well as the magnitude and direction of the signal. Data from 35 mutants are explained by a semi-empirical three-domain model in which the sensor, intervening HAMP, and catalytic domains can adopt kinase-promoting or inhibiting conformations that are in allosteric communication. The catalytic and sensor domains intrinsically favor a constitutively 'kinase-on' conformation, while the HAMP domain favors the 'off' state; when coupled, they create a bistable system responsive to physiological concentrations of Mg2+. Mutations alter signaling by locally modulating domain intrinsic equilibrium constants and interdomain couplings. Our model suggests signals transmit via interdomain allostery rather than propagation of a single concerted conformational change, explaining the diversity of signaling structural transitions observed in individual HK domains.

scholarly journals Allosteric mechanism of signal transduction in the two-component system histidine kinase PhoQ

2021 ◽  
Author(s):  
Bruk Mensa ◽  
Nicholas F Polizzi ◽  
Kathleen S Molnar ◽  
Andrew M Natale ◽  
Thomas Lemmin ◽  
...  
Keyword(s):  
Histidine Kinase ◽  
Equilibrium Constants ◽  
Kinase Domain ◽  
Domain Model ◽  
Bistable System ◽  
E Coli ◽  
Allosteric Communication ◽  
Two Component ◽  
Cytoplasmic Kinase Domain ◽  
Semi Empirical

Transmembrane signaling proteins couple extracytosolic sensors to cytosolic effectors. Here, we examine how binding of Mg2+ to the sensor domain of an E. coli two component histidine kinase (HK), PhoQ, modulates its cytoplasmic kinase domain. We use cysteine crosslinking and reporter-gene assays to simultaneously and independently probe the signaling state of PhoQ’s sensor and autokinase domains in a set of over 30 mutants. Strikingly, conservative single-site mutants distant from the sensor or catalytic site strongly influence PhoQ’s ligand-sensitivity as well as the magnitude and direction of the signal, endowing diverse signaling characteristics without need for epistasis. Data from 35 mutants are explained by a semi-empirical 3-domain model in which the sensor, intervening HAMP, and catalytic domains can adopt kinase-promoting or inhibiting conformations, that are in allosteric communication. The catalytic and sensor domains intrinsically favor a constitutively ‘kinase-on’ conformation, while the HAMP favors the ‘off’ state; when coupled, they create a bistable system responsive to physiological [Mg2+]. Mutants alter signaling by locally modulating these intrinsic equilibrium constants and couplings. Our model suggests signals transmit via interdomain allostery rather than propagation of a single concerted conformational change, explaining the diversity of signaling structural transitions observed in individual HK domains.

NMR structure of the histidine kinase domain of the E. coli osmosensor EnvZ

Nature ◽  
10.1038/23968 ◽  
1998 ◽  
Vol 396 (6706) ◽  
pp. 88-92 ◽  
Author(s):  
Toshiyuki Tanaka ◽  
Soumitra K. Saha ◽  
Chieri Tomomori ◽  
Rieko Ishima ◽  
Dingjiang Liu ◽  
...  
Keyword(s):  
Histidine Kinase ◽  
Kinase Domain ◽  
Nmr Structure ◽  
E Coli

scholarly journals Fumarate Regulation of Gene Expression in Escherichia coli by the DcuSR (dcuSR Genes) Two-Component Regulatory System

1998 ◽  
Vol 180 (20) ◽  
pp. 5421-5425 ◽  
Author(s):  
Evelyn Zientz ◽  
Johannes Bongaerts ◽  
Gottfried Unden
Keyword(s):  
Escherichia Coli ◽  
Histidine Kinase ◽  
Response Regulator ◽  
Regulation Of Gene Expression ◽  
Regulatory System ◽  
E Coli ◽  
Expression In Escherichia Coli ◽  
Two Component ◽  
Genes Encoding ◽  
Periplasmic Domain

ABSTRACT In Escherichia coli the genes encoding the anaerobic fumarate respiratory system are transcriptionally regulated by C4-dicarboxylates. The regulation is effected by a two-component regulatory system, DcuSR, consisting of a sensory histidine kinase (DcuS) and a response regulator (DcuR). DcuS and DcuR are encoded by the dcuSR genes (previouslyyjdHG) at 93.7 min on the calculated E. coli map. Inactivation of the dcuR anddcuS genes caused the loss of C4-dicarboxylate-stimulated synthesis of fumarate reductase (frdABCD genes) and of the anaerobic fumarate-succinate antiporter DcuB (dcuB gene). DcuS is predicted to contain a large periplasmic domain as the supposed site for C4-dicarboxylate sensing. Regulation by DcuR and DcuS responded to the presence of the C4-dicarboxylates fumarate, succinate, malate, aspartate, tartrate, and maleate. Since maleate is not taken up by the bacteria under these conditions, the carboxylates presumably act from without. Genes of the aerobic C4-dicarboxylate pathway encoding succinate dehydrogenase (sdhCDAB) and the aerobic succinate carrier (dctA) are only marginally or negatively regulated by the DcuSR system. The CitAB two-component regulatory system, which is highly similar to DcuSR, had no effect on C4-dicarboxylate regulation of any of the genes.

scholarly journals Succinoglycan Production by Rhizobium meliloti Is Regulated through the ExoS-ChvI Two-Component Regulatory System

1998 ◽  
Vol 180 (1) ◽  
pp. 20-26 ◽  
Author(s):  
Hai-Ping Cheng ◽  
Graham C. Walker
Keyword(s):  
Histidine Kinase ◽  
Cytoplasmic Membrane ◽  
Rhizobium Meliloti ◽  
Response Regulator ◽  
Regulatory System ◽  
Kinase Domain ◽  
Cloning And Sequencing ◽  
System A ◽  
Two Component ◽  
Close Homolog

ABSTRACT The Rhizobium meliloti exoS gene is involved in regulating the production of succinoglycan, which plays a crucial role in the establishment of the symbiosis between R. melilotiRm1021 and its host plant, alfalfa. TheexoS96::Tn5 mutation causes the upregulation of the succinoglycan biosynthetic genes, thereby resulting in the overproduction of succinoglycan. Through cloning and sequencing, we found that the exoS gene is a close homolog of theAgrobacterium tumefaciens chvG gene, which has been proposed to encode the sensor protein of the ChvG-ChvI two-component regulatory system, a member of the EnvZ-OmpR family. Further analyses revealed the existence of a newly discovered A. tumefaciens chvI homolog located just upstream of the R. meliloti exoS gene. R. meliloti ChvI may serve as the response regulator of ExoS in a two-component regulatory system. By using ExoS-specific antibodies, it was found that the ExoS protein cofractionated with membrane proteins, suggesting that it is located in the cytoplasmic membrane. By using the same antibodies, it was shown that the exoS96::Tn5 allele encodes an N-terminal truncated derivative of ExoS. The cytoplasmic histidine kinase domain of ExoS was expressed in Escherichia coli and purified, as was the R. meliloti ChvI protein. The ChvI protein autophosphorylated in the presence of acetylphosphate, and the ExoS cytoplasmic domain fragment autophosphorylated at a histidine residue in the presence of ATP. The ChvI protein was phosphorylated in the presence of ATP only when the histidine kinase domain of ExoS was also present. We propose a model for regulation of succinoglycan production by R. meliloti through the ExoS-ChvI two-component regulatory system.

scholarly journals The Anaplasma phagocytophilum PleC Histidine Kinase and PleD Diguanylate Cyclase Two-Component System and Role of Cyclic Di-GMP in Host Cell Infection

2008 ◽  
Vol 191 (3) ◽  
pp. 693-700 ◽  
Author(s):  
Tzung-Huei Lai ◽  
Yumi Kumagai ◽  
Mamoru Hyodo ◽  
Yoshihiro Hayakawa ◽  
Yasuko Rikihisa
Keyword(s):  
Histidine Kinase ◽  
Anaplasma Phagocytophilum ◽  
Kinase Domain ◽  
Etiologic Agent ◽  
Diguanylate Cyclase ◽  
Response Regulators ◽  
Cell Infection ◽  
Granulocytic Anaplasmosis ◽  
Two Component

ABSTRACT Anaplasma phagocytophilum, the etiologic agent of human granulocytic anaplasmosis (HGA), has genes predicted to encode three sensor kinases, one of which is annotated PleC, and three response regulators, one of which is PleD. Prior to this study, the roles of PleC and PleD in the obligatory intracellular parasitism of A. phagocytophilum and their biochemical activities were unknown. The present study illustrates the relevance of these factors by demonstrating that both pleC and pleD were expressed in an HGA patient. During A. phagocytophilum development in human promyelocytic HL-60 cells, PleC and PleD were synchronously upregulated at the exponential growth stage and downregulated prior to extracellular release. A recombinant PleC kinase domain (rPleCHKD) has histidine kinase activity; no activity was observed when the conserved site of phosphorylation was replaced with alanine. A recombinant PleD (rPleD) has autokinase activity using phosphorylated rPleCHKD as the phosphoryl donor but not with two other recombinant histidine kinases. rPleCHKD could not serve as the phosphoryl donor for a mutant rPleD (with a conserved aspartic acid, the site of phosphorylation, replaced by alanine) or two other A. phagocytophilum recombinant response regulators. rPleD had diguanylate cyclase activity to generate cyclic (c) di-GMP from GTP in vitro. UV cross-linking of A. phagocytophilum lysate with c-di-[32P]GMP detected an ∼47-kDa endogenous protein, presumably c-di-GMP downstream receptor. A new hydrophobic c-di-GMP derivative, 2′-O-di(tert-butyldimethylsilyl)-c-di-GMP, inhibited A. phagocytophilum infection in HL-60 cells. Our results suggest that the two-component PleC-PleD system is a diguanylate cyclase and that a c-di-GMP-receptor complex regulates A. phagocytophilum intracellular infection.

scholarly journals Comparative analysis of two paradigm bacteriophytochromes reveals opposite functionalities in two-component signaling

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Elina Multamäki ◽  
Rahul Nanekar ◽  
Dmitry Morozov ◽  
Topias Lievonen ◽  
David Golonka ◽  
...  
Keyword(s):  
Histidine Kinase ◽  
Deinococcus Radiodurans ◽  
Response Regulator ◽  
Red Light ◽  
Kinase Domain ◽  
Structural Level ◽  
Signaling Systems ◽  
Two Component ◽  
Cognate Response Regulator ◽  
Absorbing States

AbstractBacterial phytochrome photoreceptors usually belong to two-component signaling systems which transmit environmental stimuli to a response regulator through a histidine kinase domain. Phytochromes switch between red light-absorbing and far-red light-absorbing states. Despite exhibiting extensive structural responses during this transition, the model bacteriophytochrome from Deinococcus radiodurans (DrBphP) lacks detectable kinase activity. Here, we resolve this long-standing conundrum by comparatively analyzing the interactions and output activities of DrBphP and a bacteriophytochrome from Agrobacterium fabrum (Agp1). Whereas Agp1 acts as a conventional histidine kinase, we identify DrBphP as a light-sensitive phosphatase. While Agp1 binds its cognate response regulator only transiently, DrBphP does so strongly, which is rationalized at the structural level. Our data pinpoint two key residues affecting the balance between kinase and phosphatase activities, which immediately bears on photoreception and two-component signaling. The opposing output activities in two highly similar bacteriophytochromes suggest the use of light-controllable histidine kinases and phosphatases for optogenetics.

scholarly journals Characterization of the RcsC Sensor Kinase from Erwinia amylovora and Other Enterobacteria

2011 ◽  
Vol 101 (6) ◽  
pp. 710-717 ◽  
Author(s):  
Dongping Wang ◽  
Schuyler S. Korban ◽  
P. Lawrence Pusey ◽  
Youfu Zhao
Keyword(s):  
Histidine Kinase ◽  
Erwinia Amylovora ◽  
Kinase Domain ◽  
Sensor Kinase ◽  
Pear Fruit ◽  
E Coli ◽  
Pantoea Stewartii ◽  
Mutant Strains

RcsC is a hybrid sensor kinase which contains a sensor domain, a histidine kinase domain, and a receiver domain. We have previously demonstrated that, although the Erwinia amylovora rcsC mutant produces more amylovoran than the wild-type (WT) strain in vitro, the mutant remains nonpathogenic on both immature pear fruit and apple plants. In this study, we have comparatively characterized the Erwinia RcsC and its homologs from various enterobacteria. Results demonstrate that expression of the Erwinia rcsC gene suppresses amylovoran production in various amylovoran overproducing WT and mutant strains, thus suggesting the presence of a net phosphatase activity of Erwinia RcsC. Findings have also demonstrated that rcsC homologs from other enterobacteria could not rescue amylovoran production of the Erwinia rcsC mutant in vitro. However, virulence of the Erwinia rcsC mutant is partially restored by rcsC homologs from Pantoea stewartii, Yersinia pestis, and Salmonella enterica but not from Escherichia coli on apple shoots. Domain-swapping experiments have indicated that replacement of the E. coli RcsC sensor domain by those of Erwinia and Yersinia spp. partially restores virulence of the Erwinia rcsC mutant, whereas chimeric constructs containing the sensor domain of E. coli RcsC could not rescue virulence of the Erwinia rcsC mutant on apple. Interestingly, only chimeric constructs containing the histidine kinase and receiver domains of Erwinia RcsC are fully capable of rescuing amylovoran production. These results suggest that the sensor domain of RcsC may be important in regulating bacterial virulence, whereas the activity of the histidine kinase and receiver domains of Erwinia RcsC may be essential for amylovoran production in vitro.

scholarly journals Illuminating a Phytochrome Paradigm – a Light-Activated Phosphatase in Two-Component Signaling Uncovered

2020 ◽  
Author(s):  
Elina Multamäki ◽  
Rahul Nanekar ◽  
Dmitry Morozov ◽  
Topias Lievonen ◽  
David Golonka ◽  
...  
Keyword(s):  
Histidine Kinase ◽  
Deinococcus Radiodurans ◽  
Response Regulator ◽  
Red Light ◽  
Kinase Domain ◽  
Structural Level ◽  
Signaling Systems ◽  
Two Component ◽  
Cognate Response Regulator ◽  
Absorbing States

ABSTRACTBacterial phytochrome photoreceptors usually belong to two-component signaling systems which transmit environmental stimuli to a response regulator through a histidine kinase domain. Phytochromes switch between red light-absorbing and far-red light-absorbing states. Despite exhibiting extensive structural responses during this transition, the model bacteriophytochrome from Deinococcus radiodurans (DrBphP) lacks detectable kinase activity. Here, we resolve this long-standing conundrum by comparatively analyzing the interactions and output activities of DrBphP and a bacteriophytochrome from Agrobacterium fabrum (AgP1). Whereas AgP1 acts as a conventional histidine kinase, we identify DrBphP as a light-sensitive phosphatase. While AgP1 binds its cognate response regulator only transiently, DrBphP does so strongly, which is rationalized at the structural level. Our data pinpoint two key residues affecting the balance between kinase and phosphatase activities, which immediately bears on photoreception and two-component signaling. The opposing output activities in two highly similar bacteriophytochromes inform the use of light-controllable histidine kinases and phosphatases for optogenetics.

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