Design and Signaling Mechanism of Light-Regulated Histidine Kinases

Möglich, Andreas; Ayers, Rebecca A.; Moffat, Keith

doi:10.1016/j.jmb.2008.12.017

Cited by 323 publications

(381 citation statements)

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“…We speculate that in the kinase-to-phosphatase transition, the connections of the PAS cores with their flanking C-terminal ␣-helices break. The highly conserved Asp residue of the "DIT" motif, which frequently terminates PAS core domains, mediates interactions of the core with these helices (22,(39)(40)(41). In BvgS, replacement of this Asp 695 residue with Ala abolished kinase activity, a finding consistent with this scenario (28).…”

Section: Mechanical Signal Transmission By Bvgs Linkermentioning

confidence: 59%

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Conformational Changes of an Interdomain Linker Mediate Mechanical Signal Transmission in Sensor Kinase BvgS

et al. 2017

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The whooping cough agent, Bordetella pertussis, controls the expression of its large virulence regulon in a coordinated manner through the two-component system BvgAS. BvgS is a dimeric, multidomain sensor kinase. Each monomer comprises, in succession, tandem periplasmic Venus flytrap (VFT) domains, a transmembrane segment, a cytoplasmic Per-Arnt-Sim (PAS) domain, a kinase module, and additional phosphorelay domains. BvgS shifts between kinase and phosphatase modes of activity in response to chemical modulators that modify the clamshell motions of the VFT domains. We have shown previously that this regulation involves a shift between distinct states of conformation and dynamics of the two-helix coiled-coil linker preceding the enzymatic module. In this work, we determined the mechanism of signal transduction across the membrane via a first linker, which connects the VFT and PAS domains of BvgS, using extensive cysteine cross-linking analyses and other approaches. Modulator perception by the periplasmic domains appears to trigger a small, symmetrical motion of the transmembrane segments toward the periplasm, causing rearrangements of the noncanonical cytoplasmic coiled coil that follows. As a consequence, the interface of the PAS domains is modified, which affects the second linker and eventually causes the shift of enzymatic activity. The major features of this first linker are well conserved among BvgS homologs, indicating that the mechanism of signal transduction unveiled here is likely to be generally relevant for this family of sensor kinases.IMPORTANCE Bordetella pertussis produces virulence factors coordinately regulated by the two-component system BvgAS. BvgS is a sensor kinase, and BvgA is a response regulator that activates gene transcription when phosphorylated by BvgS. Sensor kinases homologous to BvgS are also found in other pathogens. Our goal is to decipher the mechanisms of BvgS signaling, since these sensor kinases may represent new targets for antibacterial agents. Signal perception by the sensor domains of BvgS triggers small motions of the helical linker region underneath. The protein domain that follows this linker undergoes a large conformational change that amplifies the initial signal, causing a shift of activity from kinase to phosphatase. Because BvgS homologs harbor similar regions, these signaling mechanisms are likely to apply generally to that family of sensor kinases.

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Section: Mechanical Signal Transmission By Bvgs Linkermentioning

confidence: 59%

“…Signal transduction between the perception and the enzymatic domains of sensor kinases generally involves combinations of rotation, scissor, or piston movements of coiled-coil regions (40)(41)(42)(43)(44)(45)(46)(47)(48)(49). A simple model is that a sensor kinase populates two thermodynamically stable structural states.…”

Section: Discussionmentioning

confidence: 99%

Conformational Changes of an Interdomain Linker Mediate Mechanical Signal Transmission in Sensor Kinase BvgS

et al. 2017

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“…Interestingly, many histidine kinases that have a PAS domain as signal input domain, show conservation of the connecting region between the PAS domain and the J␣-helix (47). This conserved sequence signature ( 125 DIT 127 ) is also present in YtvA (Fig.…”

Section: Discussionmentioning

confidence: 91%

“…Interdomain Signal Transmission in YtvA-A recent study of engineered hybrid proteins, composed of the LOV domain of YtvA and the histidine kinase domain of FixL and connected via J␣-type helices of different lengths, has revealed that signals are transmitted from the sensory LOV domain to the histidine kinase domain via a 40 -60 o rotational movement within the ␣-helical coiled-coil linker (47). Therefore, these hybrid proteins act as light-triggered rotary switches (7,47).…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, by examining an alignment of LOV-STAS proteins, it is possible to recognize the typical hydrophobic pattern (Fig. 4) that defines a coiled-coil structure, just as in the J␣-helix between several PAS domains and their cognate histidine kinase domain (47). Based on the conserved sequences pattern between the above mentioned hybrid protein and YtvA-J␣, we anticipate that the STAS domain is activated via a coiled-coil structure of dimeric YtvA, in a way similar to what has been described for PAS domain containing histidine kinases.…”

Section: Discussionmentioning

confidence: 99%

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In Vivo Mutational Analysis of YtvA from Bacillus subtilis

Ávila-Pérez¹,

Vreede

Tang

et al. 2009

Journal of Biological Chemistry

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LOV2 domains (1), members of the superfamily of PAS domains (2, 3), are abundant in all domains of life and were first identified in plant phototropins (4). These photoreceptors regulate stomatal opening, phototropism, etc. and contain two N-terminal LOV domains that confer light regulation on the C-terminal Ser/Thr kinase domain (4). They also occur in bacteria, in which YtvA from Bacillus subtilis has been best characterized (for a review, see e.g. Ref. 5). Its N-terminal LOV domain binds FMN and shows the typical LOV photochemistry (6, 7): covalent adduct formation between a cysteine and the FMN chromophore. A linker helix, denoted J␣ (7), connects the LOV domain to a STAS domain. The latter domain is present in many regulators of the general stress response of B. subtilis (8,9). Stress via the addition of salt or ethanol (for a review, see Ref.10) and blue light (11, 12) activates the general stress response via the environmental pathway, which integrates various signals via a large multiprotein complex, called the stressosome (13,14). YtvA, which mediates light activation of B (11, 12, 15), co-purifies with other STAS domain proteins in the stressosomes (16). When cells are stressed, STAS domains of several stressosome proteins (e.g. RsbS and RsbR) are phosphorylated by another intrinsic stressosome component, the serine/threonine kinase RsbT (9,14,17,18). Next, RsbT is released from the complex to trigger RsbU, a protein phosphatase, thus (indirectly) activating B (19). Phosphorylation of YtvA, however, has never been detected. Rather, it has been demonstrated in vitro that YtvA shows light-dependent GTP binding, presumably at its NTP-binding site in the STAS domain (20).Little is known about the mechanism of signal transmission in and by YtvA, except that in the C62A mutant, photochemistry in vitro (12) and light activation of B in vivo (12, 15) are abolished. More detailed information is available for LOV domains of phototropins. A conserved glutamine, which is in hydrogen-bonding contact with the isoalloxazine ring of FMN, rotates its side chain by 180 o upon covalent adduct formation (21). Replacement of this residue by leucine in the LOV2 domain of Phy3 from Adiantum results in a considerable reduction of the light-induced structural change (22). The corresponding mutation in phototropin 1 from Arabidopsis impairs autophosphorylation activity (23). The signal generated in the LOV2 domain is transmitted to the downstream kinase domain of phototropin 1 of Avena sativa through disruption of the interaction between its central ␤-sheet and the C-terminal linker region, the J␣-helix (24).Here, we study the mechanism of activation of YtvA in vivo, i.e. light-induced activation of the B response, with site-directed mutagenesis. We focus on three regions of the protein, the flavinbinding pocket, the ␤-sheet of the LOV domain, and the GTPbinding site, and on potential phosphorylation sites of the STAS domain. We demonstrate that light-activated GTP binding is crucial for functional YtvA. A computational approac...

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Dynamics and Kinetics in Structural Biology

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Design and Signaling Mechanism of Light-Regulated Histidine Kinases

Cited by 323 publications

References 50 publications

Conformational Changes of an Interdomain Linker Mediate Mechanical Signal Transmission in Sensor Kinase BvgS

Conformational Changes of an Interdomain Linker Mediate Mechanical Signal Transmission in Sensor Kinase BvgS

In Vivo Mutational Analysis of YtvA from Bacillus subtilis

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