Segmental Helical Motions and Dynamical Asymmetry Modulate Histidine Kinase Autophosphorylation

Mechaly, A.E.; Sassoon, Nathalie; Betton, Jean‐Michel; Alzari, Pedro M.

doi:10.1371/journal.pbio.1001776

Cited by 111 publications

(214 citation statements)

References 66 publications

(94 reference statements)

Supporting

Mentioning

171

Contrasting

Unclassified

Order By: Relevance

“…6B). This model is supported by several dimeric HK structures that show that the CA domain can adopt different arrangements with respect to the DHp domain, demonstrating the plasticity of this interface (28,36). Although signaling relies on direct contact between the LOV β-sheet and the DHpL domain for EL346, we envision that, in dimeric HKs, it depends on sensors acting at a distance, modulating the movement of DHp helices from the two protein chains relative to each other.…”

Section: Discussionmentioning

confidence: 66%

“…R143A and E246A mutations diminished autokinase activity to levels below those measurable by our assays. Based on the recently reported structures of the autophosphorylating Michaelis complex (20,28), we predict that Glu246 acts as the catalytic base whereas Arg143 helps position it through hydrogen bonding in the active state, making both residues essential for catalysis. Although we cannot precisely determine what specific changes are induced in the DHpL domain without detailed structural information on activated EL346, we can attempt to mimic the active site by superposing the individual DHpL and CA domains from the EL346FL structure onto the autophosphorylationcompetent chain of the Enz chim structure (Fig.…”

Section: Mutations At the Domain Interfaces Affect Signaling And Automentioning

confidence: 98%

See 1 more Smart Citation

Full-length structure of a monomeric histidine kinase reveals basis for sensory regulation

Rivera-Cancel

Tomchick

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

Although histidine kinases (HKs) are critical sensors of external stimuli in prokaryotes, the mechanisms by which their sensor domains control enzymatic activity remain unclear. Here, we report the full-length structure of a blue light-activated HK from Erythrobacter litoralis HTCC2594 (EL346) and the results of biochemical and biophysical studies that explain how it is activated by light. Contrary to the standard view that signaling occurs within HK dimers, EL346 functions as a monomer. Its structure reveals that the light-oxygen-voltage (LOV) sensor domain both controls kinase activity and prevents dimerization by binding one side of a dimerization/histidine phosphotransfer-like (DHpL) domain. The DHpL domain also contacts the catalytic/ATP-binding (CA) domain, keeping EL346 in an inhibited conformation in the dark. Upon light stimulation, interdomain interactions weaken to facilitate activation. Our data suggest that the LOV domain controls kinase activity by affecting the stability of the DHpL/CA interface, releasing the CA domain from an inhibited conformation upon photoactivation. We suggest parallels between EL346 and dimeric HKs, with sensor-induced movements in the DHp similarly remodeling the DHp/CA interface as part of activation.two-component system | cell signaling | histidine kinase | photosensory | regulation

show abstract

Section: Discussionmentioning

confidence: 66%

Section: Mutations At the Domain Interfaces Affect Signaling And Automentioning

confidence: 98%

Full-length structure of a monomeric histidine kinase reveals basis for sensory regulation

Rivera-Cancel

Tomchick

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

show abstract

“…The activity of histidine kinases is modulated by environmental cues through signal-induced conformational changes (Bhate et al 2015;Mechaly et al 2014;Wang et al 2013). Chemical cross-linking and size-exclusion chromatography were utilised in order to understand the effect of salt stress on Fig.…”

Section: Discussionmentioning

confidence: 99%

Classical macroautophagy in Lobivia rauschii (Cactaceae) and possible plastidial autophagy in Tillandsia albida (Bromeliaceae) tapetum cells

2013

View full text Add to dashboard Cite

Two-component signal transduction systems (TCSs) consist of sensor histidine kinases and response regulators. TCSs mediate adaptation to environmental changes in bacteria, plants, fungi and protists. Histidine kinase 2 (Hik2) is a sensor histidine kinase found in all known cyanobacteria and as chloroplast sensor kinase in eukaryotic algae and plants. Sodium ions have been shown to inhibit the autophosphorylation activity of Hik2 that precedes phosphoryl transfer to response regulators, but the mechanism of inhibition has not been determined. We report on the mechanism of Hik2 activation and inactivation probed by chemical crosslinking and size exclusion chromatography together with direct visualisation of the kinase using negative-stain transmission electron microscopy of single particles. We show that the functional form of Hik2 is a higher-order oligomer such as a hexamer or octamer. Increased NaCl concentration converts the active hexamer into an inactive tetramer. The action of NaCl appears to be confined to the Hik2 kinase domain.

show abstract

“…To probe the contribution of CroS kinase activity toward CroR phosphorylation and cephalosporin resistance, we introduced substitutions at the conserved histidine predicted to be the site of autophosphorylation (H172) or the adjacent acidic residue (D173) which is critical for HK autophosphorylation in other TCS kinases (9,10,45). Phos-tag analysis demonstrated that the CroS H172A variant resulted in some CroR-P that can be detected in a culture without CroRS stimulation, while the CroS D173A variant cannot produce CroR-P even upon stimulation of CroS (Fig.…”

Section: Analysis Of Gene Expression By Quantitative Reverse Transcrimentioning

confidence: 99%

Functional Dissection of the CroRS Two-Component System Required for Resistance to Cell Wall Stressors in Enterococcus faecalis

Kellogg

Kristich

2016

J Bacteriol

View full text Add to dashboard Cite

Bacteria use two-component signal transduction systems (TCSs) to sense and respond to environmental changes via a conserved phosphorelay between a sensor histidine kinase and its cognate response regulator. The opportunistic pathogen Enterococcus faecalis utilizes a TCS comprised of the histidine kinase CroS and the response regulator CroR to mediate resistance to cell wall stresses such as cephalosporin antibiotics, but the molecular details by which CroRS promotes cephalosporin resistance have not been elucidated. Here, we analyzed mutants of E. faecalis carrying substitutions in CroR and CroS to demonstrate that phosphorylated CroR drives resistance to cephalosporins, and that CroS exhibits kinase and phosphatase activities to control the level of CroR phosphorylation in vivo. Deletion of croS in various lineages of E. faecalis revealed a CroS-independent mechanism for CroR phosphorylation and led to the identification of a noncognate histidine kinase capable of influencing CroR (encoded by OG1RF_12162; here called cisS). Further analysis of this TCS network revealed that both systems respond to cell wall stress. IMPORTANCETCSs allow bacteria to sense and respond to many different environmental conditions. The opportunistic pathogen Enterococcus faecalis utilizes the CroRS TCS to mediate resistance to cell wall stresses, including clinically relevant antibiotics such as cephalosporins and glycopeptides. In this study, we use genetic and biochemical means to investigate the relationship between CroRS signaling and cephalosporin resistance in E. faecalis cells. Through this, we uncovered a signaling network formed between the CroRS TCS and a previously uncharacterized TCS that also responds to cell wall stress. This study provides mechanistic insights into CroRS signaling and cephalosporin resistance in E. faecalis. Although Enterococcus faecalis is a normal commensal of the gut microbiome, it poses a serious risk to humans as an opportunistic pathogen. A major risk factor for the pathogenic transition of enterococci is therapy with broad-spectrum cephalosporin antibiotics. Cephalosporins are commonly used to treat bacterial infections; however, nearly all enterococcal isolates are resistant to these compounds. This intrinsic resistance allows for the expansion of resident enterococcal populations during cephalosporin treatment, which is thought to promote dissemination from the gastrointestinal tract, enabling enterococci to establish infections (1, 2). With the emergence of multidrug-resistant isolates of E. faecalis, few options for treating infections are available. Therefore, understanding the mechanisms that support cephalosporin resistance in E. faecalis is crucial for designing new therapies that limit growth during cephalosporin treatment or that can be used as adjuvants with cephalosporins to treat E. faecalis infections. An essential determinant of cephalosporin resistance in E. faecalis is the CroRS two-component system (TCS), consisting of CroS (a transmembrane sensor-histidine kinase) and ...

show abstract

Segmental Helical Motions and Dynamical Asymmetry Modulate Histidine Kinase Autophosphorylation

Abstract: Step-by-step structures of a prototypical E. coli histidine kinase reveal how external stimuli drive helical bending motions to control asymmetric movements of the catalytic domains.

Cited by 111 publications

References 66 publications

Full-length structure of a monomeric histidine kinase reveals basis for sensory regulation

Full-length structure of a monomeric histidine kinase reveals basis for sensory regulation

Classical macroautophagy in Lobivia rauschii (Cactaceae) and possible plastidial autophagy in Tillandsia albida (Bromeliaceae) tapetum cells

Functional Dissection of the CroRS Two-Component System Required for Resistance to Cell Wall Stressors in Enterococcus faecalis

Contact Info

Product

Resources

About