Balance between Coiled-Coil Stability and Dynamics Regulates Activity of BvgS Sensor Kinase in
            <i>Bordetella</i>

Lesne, E.; Krammer, Eva-Maria; Dupré, Elian; Locht, Camille; Lensink, Marc F.; Antoine, Rudy; Jacob‐Dubuisson, Françoise

doi:10.1128/mbio.02089-15

Cited by 22 publications

(78 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, we measured the proportions of spontaneous in vivo intermonomer disulfide (S-S) bond-mediated cross-linking in bacteria grown under default (i.e., kinase-promoting) or modulating (i.e., phosphatase-promoting) conditions. These substitutions were introduced into a full-length version of BvgS called BvgS fl , in which two naturally occurring Cys 607 and Cys 881 residues were replaced by Ala and Ser, respectively (29). Remarkably, high proportions of S-S cross-linking were detected at all three positions, with dimer-to-monomer ratios of approximately 70% under both growth conditions (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To determine the organization of the hydrophobic segment of linker 1, Cys-scanning S-S cross-linking analyses were performed in Escherichia coli using a truncated BvgS variant called BvgS t . BvgS t is devoid of the receiver and HPt domains, and it harbors mutations of its natural Cys residues as described above, as well as a C-terminal 6-His tag for immune detection (29). The corresponding BvgS fl variants were also constructed to determine BvgS activity in B. pertussis.…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, dimer formation globally decreased when nicotinate was added to the bacteria, arguing in favor of a change of interface. This change is likely to have a direct effect on the downstream linker 2, whose conformation and dynamics eventually determine the mode of activity of the enzymatic moiety (29).…”

Section: S7bmentioning

confidence: 99%

“…Our earlier studies have indicated the remarkable conservation of the BvgS PAS domain among Bordetella isolates and have shown the importance of the PAS core fold for the regulation of BvgS activity (14,28). We have proposed that the PAS domain serves as a toggle switch (29).…”

mentioning

confidence: 99%

“…The kinase mode is characterized by considerable rotational dynamics in this region, whereas in the phosphatase mode, linker 2 adopts its stable, hydrophobic coiled-coil interface (29). The manner in which the dynamics of VFT domains are transmitted to linker 2, and in particular the mechanism by which the intervening linker 1 and PAS domains of BvgS mediate this signaling, is the focus of this work.…”

mentioning

confidence: 99%

See 4 more Smart Citations

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

et al. 2017

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: S7bmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

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

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

Regulatory Systems: Two‐Component

Raivio

2019

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Two‐component signal transduction (TCST) systems constitute a large class of regulatory proteins that function as signal transducers. Each system comprises a sensor or histidine kinase (HK) and an effector or response regulator (RR), which communicate through a conserved set of phosphotransfer reactions to effect adaptive changes in response to specific environmental signals. HKs and RRs are modular in nature, with variable sensory and output structures appended to the conserved domains that facilitate phosphotransfer mediated signal transduction. Input signals trigger successive conformational changes in domains and protein:protein interactions that alter phosphotransfer, and ultimately an output response mediated by the phosphorylated RR. TCST systems are abundant in bacteria and many microbes utilise multiple TCST pathways to sense and respond to a plethora of environmental and physiological changes. Specificity between cognate HKs and RRs is largely maintained through co‐evolving residues at a conserved interface where the RR docks on the HK. TCST systems are integrated into cellular signalling networks and interact with macromolecules and proteins that connect them to salient regulatory pathways and cellular functions. The current state of knowledge around TCST systems will be summarised, emphasising findings published since the first version of this article in 2006. Key Concepts A prototypical two‐component system is made up of a membrane‐bound sensory histidine kinase that senses a unique environmental or cellular parameter and a cytoplasmic response regulator that controls an adaptive response. HKs and RRs communicate through phosphotransfer reactions that are mediated by conserved domains; signal sensing alters the ratio of HK kinase to phosphatase activity to change the function of the RR by altering its phosphorylation status. HKs and RRs are organised in a modular fashion; a variety of sensing domains can be appended to the HK enzymatic module while diverse output domains with DNA binding, RNA binding, enzymatic activity or protein binding functions are often fused to the C‐terminal end of the response regulator phosphorylated receiver (REC) domain. The HK is a dimer containing a catalytic domain composed of two DHp and CA domains. The DHp domain consists of a d imer of two alpha helices connected by a flexible linker that makes a four‐helical bundle and contains the conserved h istidine that is the site of auto p hosphorylation. The CA domain resembles other ATP‐binding folds and is the enzymatic portion of the HK. The RR consists at its N‐terminus of the conserved receiver (REC) domain, which consists of a five‐stranded beta sheet surrounded by alpha helices and contains the conserved aspartate that is the site of phosphorylation. Signals are detected through conformational changes in a sensory domain that are propagated through some combination of rotational, piston and order to disorder transitions by alpha‐helical transduction elements to the cytoplasmic enzymatic domain of the HK. These movements lead to alterations in HK dimer symmetry that dictate kinase or phosphatase activity. In the inactive state, the cytoplasmic DHp and CA domains of the HK are organised in a symmetrical fashion with the CA domain juxtaposed against the membrane‐proximal end of the DHp four‐helical bundle. The RR REC domain can interact with the inactive HK DHp four‐helical bundle at a more membrane‐distal location in an orientation that would facilitate dephosphorylation of the aspartate. The active, kinase state of the HK is asymmetrical due to a bend or kink in the DHp domain that leads to one CA domain adopting a looser association that positions it well for phosphorylation of a histidine residue. A single RR REC domain can bind near the other CA domain, which is more closely associated with the DHp domain, in a conformation supporting phosphorylation of the aspartate residue utilising the phosphorylated histidine as a substrate. Repeated cycles of DHp domain bending and RR REC domain binding are hypothesised to support successive rounds of HK autophosphorylation and RR phosphorylation in response to signal inputs received from the sensory domain. HKs and RRs regulate and interact with other macromolecules and proteins to connect signalling pathways, coordinate their activities and sense environmental parameters and changes to cellular physiology.

show abstract

CCBuilder 2.0: Powerful and accessible coiled‐coil modeling

2017

View full text Add to dashboard Cite

The increased availability of user‐friendly and accessible computational tools for biomolecular modeling would expand the reach and application of biomolecular engineering and design. For protein modeling, one key challenge is to reduce the complexities of 3D protein folds to sets of parametric equations that nonetheless capture the salient features of these structures accurately. At present, this is possible for a subset of proteins, namely, repeat proteins. The α‐helical coiled coil provides one such example, which represents ≈ 3–5% of all known protein‐encoding regions of DNA. Coiled coils are bundles of α helices that can be described by a small set of structural parameters. Here we describe how this parametric description can be implemented in an easy‐to‐use web application, called CCBuilder 2.0, for modeling and optimizing both α‐helical coiled coils and polyproline‐based collagen triple helices. This has many applications from providing models to aid molecular replacement for X‐ray crystallography, in silico model building and engineering of natural and designed protein assemblies, and through to the creation of completely de novo “dark matter” protein structures. CCBuilder 2.0 is available as a web‐based application, the code for which is open‐source and can be downloaded freely. http://coiledcoils.chm.bris.ac.uk/ccbuilder2.Lay SummaryWe have created CCBuilder 2.0, an easy to use web‐based application that can model structures for a whole class of proteins, the α‐helical coiled coil, which is estimated to account for 3–5% of all proteins in nature. CCBuilder 2.0 will be of use to a large number of protein scientists engaged in fundamental studies, such as protein structure determination, through to more‐applied research including designing and engineering novel proteins that have potential applications in biotechnology.

show abstract

Balance between Coiled-Coil Stability and Dynamics Regulates Activity of BvgS Sensor Kinase in Bordetella

Cited by 22 publications

References 45 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

Regulatory Systems: Two‐Component

CCBuilder 2.0: Powerful and accessible coiled‐coil modeling

Contact Info

Product

Resources

About