Crystal structures of the CO  and NO Bound DosS GAF-A domain and implications for DosS signaling in Mycobacterium tuberculosis

Madrona, Y.; Waddling, C.A.; Montellano, Paul R. Ortiz de

doi:10.1016/j.abb.2016.10.005

Cited by 14 publications

(21 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A). During the past 10 years, a series of interesting studies on DevS and DosT have been carried out elucidating some key residues involved in signal transduction, robust oxidative processes, NO deoxygenation, and the high‐resolution X‐ray structures of the isolated domains, among others .…”

Section: Introductionmentioning

confidence: 99%

“…Currently, tuberculosis kills over 1.4 million people every year around the world and exists in a latent state in one-third of the human population [3]. These alarming numbers highlight this transduction, robust oxidative processes, NO deoxygenation, and the high-resolution X-ray structures of the isolated domains, among others [9][10][11][12][13][14][15][16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Oxygen triggers signal transduction in the DevS (DosS) sensor of Mycobacterium tuberculosis by modulating the quaternary structure

et al. 2019

View full text Add to dashboard Cite

A major challenge to the control and eventual eradication of Mycobacterium tuberculosis infection is this pathogen's prolonged dormancy. The heme‐based oxygen sensor protein DevS (DosS) plays a key role in this phenomenon, because it is a major activator of the transcription factor DevR. When DevS is active, its histidine protein kinase region is ON and it phosphorylates and activates DevR, which can induce the transcription of the dormancy regulon genes. Here, we have investigated the mechanism by which the ligation of molecular oxygen to a heme‐binding domain in DevS switches OFF its histidine protein kinase region. To shed light on the oligomerization states of this protein and possible protein‐surfaces of interaction, we used analytical gel filtration, together with dynamic light scattering, fluorescence spectroscopy and chemical crosslinking. We found that DevS exists as three major species: an octamer, a tetramer and a dimer. These three states were observed for the concentration range between 0.5 and 20 μm DevS, but not below 0.1 μm. Levels of DevS in M. tuberculosis are expected to range from 5 to 26 μm. When this histidine protein kinase was OFF, the DevS was mainly tetrameric and dimeric; by contrast, when the kinase was ON, the protein was predominantly octameric. The changes in quaternary structure were rapid upon binding to the physiological signal. This finding represents a novel strategy for switching the activity of a two‐component heme‐based sensor. An enhanced understanding of this process might potentially lead to the design of novel regulatory agents that target the multimer interfaces for treatment of latent tuberculosis.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Oxygen triggers signal transduction in the DevS (DosS) sensor of Mycobacterium tuberculosis by modulating the quaternary structure

et al. 2019

View full text Add to dashboard Cite

show abstract

“…The X‐ray crystal structures for the HD of DevS were solved for unbonded and bonded states to CO, NO, and CN − . While met‐DevS and DevS‐CN are kinase inactive, all ferrous states, unbounded and bounded to CO and NO, are kinase active (Fig.…”

Section: Discussionmentioning

confidence: 99%

A spectroelectrochemical investigation of the heme‐based sensor DevS from Mycobacterium tuberculosis: a redox versus oxygen sensor

et al. 2019

View full text Add to dashboard Cite

Tuberculosis is one of the oldest known infectious diseases, responsible for millions of deaths annually around the world. The ability of Mycobacterium tuberculosis (Mtb) to enter into a dormant state has been considered integral to the success of this bacterium as a human pathogen. One of the key systems involved in regulating the entrance into dormancy is the differentially expressed in virulent strain sensor protein (DevS) [(dormancy survival sensor protein (DosS)]. However, the physiological signal for DevS has remained unclear since it was first shown to be a heme‐based sensor with conflicting reports on whether it is a redox or an oxygen sensor. To address this question and provide a better understanding of the electronic properties of this protein, we present here, for the first time, a series of spectroelectrochemistry measurements of the full‐length holo DevS in anaerobic conditions as well as bound to CO, NO, imidazole (Imz), cyanide, and O2. An interesting feature of this protein is its ability to bind Imz even in the ferrous state, implying small‐molecule analogues could be designed as potential regulators. Nonetheless, a midpoint potential (Em) value of +10 mV [vs normal hydrogen electrode (NHE)] for DevS as measured under anaerobic conditions is much higher than the expected cytosolic potential for Mtb or even within stimulated macrophages (~ −270 mV vs NHE), indicating this sensor works in a reduced ferrous state. These data, along with the high oxygen affinity and very slow auto‐oxidation rate of DevS, provides evidence that it is not a redox sensor. Overall, this study validates the biological function of DevS as an oxygen sensor directly involved in the dormancy/latency of Mtb.

show abstract

“…In addition to acting as an alternative energy source at low concentrations and respiratory toxin at high concentrations, CO has been shown to influence gene expression in mycobacteria, via the two-component DosS/DosR system (8, 26). The sensor histidine kinase DosS is a hemoprotein that activates the transcriptional regulator DosR via phosphorylation in response to low oxygen, low redox state or via binding of ligands to its heme functional group (27). The DosS/DosR regulator has been most thoroughly characterized in M. tuberculosis , which possesses an additional sensor kinase designated DosT that acts synergistically with DosS to modulate DosR function (28).…”

Section: Introductionmentioning

confidence: 99%

“…The DosS/DosR regulator has been most thoroughly characterized in M. tuberculosis , which possesses an additional sensor kinase designated DosT that acts synergistically with DosS to modulate DosR function (28). In M. tuberculosis , the dos regulon contains 48 genes and contributes to survival during hypoxia-induced dormancy (27, 29, 30). While the Dos response plays a role in adaption to hypoxia and for resistance to respiratory inhibition by NO, the physiological role of its activation in response to CO in M. tuberculosis has not been determined (8, 26, 30).…”

Section: Introductionmentioning

confidence: 99%

Mycobacteria tolerate carbon monoxide by remodelling their respiratory chain

Bayly

Cordero

Schittenhelm

et al. 2020

Preprint

View full text Add to dashboard Cite

18Carbon monoxide (CO) is a gas infamous for its acute toxicity. The toxicity of CO 19 predominantly stems from its tendency to form carbonyl complexes with transition 20 metals, thus inhibiting the heme-prosthetic groups of proteins, including the terminal 21 oxidases of the respiratory chain. While CO has been proposed as an antibacterial 22 agent, the evidence supporting its toxicity towards bacteria is equivocal, and its 23 cellular targets remain poorly defined. In this work, we investigate the physiological 24 response of mycobacteria to CO. We show that Mycobacterium smegmatis is highly 25 resistant to the toxic effects of CO, exhibiting normal growth parameters when 26 cultured in its presence. We profiled the proteome of M. smegmatis during growth in 27 CO, identifying strong induction of cytochrome bd oxidase and members of the dos 28 regulon, but relatively few other changes. We show that the activity of cytochrome bd 29 oxidase is resistant to CO, whereas cytochrome bcc-aa 3 oxidase is strongly inhibited 30 by this gas. Consistent with these findings, growth analysis shows that M. smegmatis 31 lacking cytochrome bd oxidase displays a significant growth defect in the presence 32 of CO, while induction of the dos regulon appears to be unimportant for adaption to 33 CO. Altogether, our findings suggest that M. smegmatis has considerable resistance 34 to CO and benefits from respiratory flexibility to withstand its inhibitory effects. 35 36 Importance 37 Carbon monoxide has an infamous reputation as a toxic gas and it has been 38 suggested that it has potential as an antibacterial agent. Despite this, the means by 39 which bacteria resist its toxic effects are not well understood. In this study we 40 determine the physiological response of Mycobacterium smegmatis to growth in CO. 41 We show for the first time that the cytochrome bd oxidase is inherently resistant to 42 CO and is deployed by M. smegmatis to tolerate the presence of this gas. Further, 43we show that aside from this remodelling of its respiratory chain, M. smegmatis 44 makes few other functional changes to its proteome, suggesting it has a high level of 45 inherent resistance to CO. 46 47

show abstract

Crystal structures of the CO and NO Bound DosS GAF-A domain and implications for DosS signaling in Mycobacterium tuberculosis

Cited by 14 publications

References 34 publications

Oxygen triggers signal transduction in the DevS (DosS) sensor of Mycobacterium tuberculosis by modulating the quaternary structure

Oxygen triggers signal transduction in the DevS (DosS) sensor of Mycobacterium tuberculosis by modulating the quaternary structure

A spectroelectrochemical investigation of the heme‐based sensor DevS from Mycobacterium tuberculosis: a redox versus oxygen sensor

Mycobacteria tolerate carbon monoxide by remodelling their respiratory chain

Contact Info

Product

Resources

About