The importance of conserved amino acids in heme-based globin-coupled diguanylate cyclases

Wan, Xuehua; Saito, Jennifer A.; Newhouse, James S.; Hou, Shaobin; Alam, Maqsudul

doi:10.1371/journal.pone.0182782

Cited by 4 publications

(4 citation statements)

References 49 publications

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“…The similar secondary structures ( Figure 4) and ligand-binding abilities (Supplementary Figure S3) of the PccGCS WT and π-helix mutant (H237A/K238A construct) but major differences in DGC activities and oligomerization ( Figure 4) suggest significant changes in quaternary structure and loss of π-helix interactions leading to interrupted signal transduction. In two previous studies by Wan and colleagues, mutation of the conserved histidine and lysine residues within EcDosC and BpeGReg led to inactive phenotypes for the two enzymes [9,10], supporting our data highlighting this region as being critical for enzyme activity of GCS proteins.…”

Section: Discussionsupporting

confidence: 90%

“…Oxygen (O 2 ) levels have been demonstrated to regulate biofilm formation and to affect virulence, making it an important external signal, likely due to its importance in metabolism [9][10][11]. Globin coupled sensor (GCS) proteins utilize a heme-containing globin domain to sense O 2 levels and transmit the ligand-binding signal through a linking middle domain to an output domain.…”

Section: Introductionmentioning

confidence: 99%

“…First, chemical cross-linking followed by protease digestion and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was used to identify interactions between domains within the two DGC-containing GCS proteins. Next, influenced by the cross-linking results and previous BpeGReg mutation studies by Wan and others [9,10], conserved histidine and lysine residues within a π-helical region of the middle domain were mutated in PccGCS, and this mutant was compared biochemically and biophysically to the wild-type form to show that the π-helix mutations do affect DGC activity. Furthermore, enzyme kinetic assays utilizing isolated diguanylate cyclase and globin domains of PccGCS revealed enhanced activity due to specific interactions of the cognate globin domain.…”

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confidence: 99%

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π-Helix controls activity of oxygen-sensing diguanylate cyclases

Walker

Potter

et al. 2020

Bioscience Reports

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The ability of organisms to sense and adapt to oxygen levels in their environment leads to changes in cellular phenotypes, including biofilm formation and virulence. Globin coupled sensors (GCSs) are a family of heme proteins that regulate diverse functions in response to O 2 levels, including modulating synthesis of cyclic dimeric guanosine monophosphate (c-di-GMP), a bacterial second messenger that regulates biofilm formation. While GCS proteins have been demonstrated to regulate O 2 -dependent pathways, the mechanism by which the O 2 binding event is transmitted from the globin domain to the cyclase domain is unknown. Using chemical cross-linking and subsequent liquid chromatography-tandem mass spectrometry, diguanylate cyclase (DGC)-containing GCS proteins from Bordetella pertussis (BpeGReg) and Pectobacterium carotovorum (PccGCS) have been demonstrated to form direct interactions between the globin domain and a middle domain π-helix. Additionally, mutation of the π-helix caused major changes in oligomerization and loss of DGC activity. Furthermore, results from assays with isolated globin and DGC domains found that DGC activity is affected by the cognate globin domain, indicating unique interactions between output domain and cognate globin sensor. Based on these studies a compact GCS structure, which depends on the middle domain π-helix for orienting the three domains, is needed for DGC activity and allows for direct sensor domain interactions with both middle and output domains to transmit the O 2 binding signal. The insights from the present study improve our understanding of DGC regulation and provide insight into GCS signaling that may lead to the ability to rationally control O 2 -dependent GCS activity.identified in numerous bacteria with a variety of output domains that are regulated by the O 2 -binding signal, including methyl accepting chemotaxis protein (MCP), kinase, and diguanylate cyclase (DGC) [12,13]. In Bacillus subtilis, the MCP-containing GCS regulates aerotaxis, allowing the organism to move to its preferred location within an O 2 gradient [14]. In contrast, the DGC-containing GCS from the plant pathogen Pectobacterium carotovorum (PccGCS) regulates O 2 -dependent motility, virulence factor production, and rotting within a plant host, highlighting the importance of O 2 sensing and GCS proteins in controlling bacterial phenotypes [11].Within the GCS protein family, the most common effect of O 2 binding to the heme of the globin domain is an increase in the activity of the output domain [13]. This increase in GCS enzymatic activity often requires oligomerization of GCS monomers to yield activity of the output domains; GCS proteins with diguanylate cyclase, histidine kinase, and adenylate cylcase output domains have been shown to function as homodimers and higher order oligomers [15][16][17][18][19][20]. Because of the multi-domain organization and oligomerization of GCS proteins, questions have arisen regarding the overall structure and path of signal transduction within the protein family. ...

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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π-Helix controls activity of oxygen-sensing diguanylate cyclases

Walker

Potter

et al. 2020

Bioscience Reports

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“…Based on in vitro reactivity, however, it is possible that GCSs could function in CO ( 76 , 78 , 79 ) or HCN sensing ( 76 , 78 , 79 ), although a physiological role for these latter functions has not been established. Coordination of O 2 in the binding pocket by Fe(II)-heme and distal threonine and tyrosine residues ( 80 ) causes the α-helices to rearrange ( 75 , 76 , 78 , 79 , 81 ), and this change is transmitted to the adjacent output domain through a coiled-coil region at the C terminus of the GCS ( 77 ).…”

Section: Modular Sensory and Receiver Domains Found In Dgcs And C-di-...mentioning

confidence: 99%

Sensory Perception in Bacterial Cyclic Diguanylate Signal Transduction

et al. 2022

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Cyclic diguanylate (c-di-GMP) signal transduction systems provide bacteria the ability to sense changing cell status or environmental conditions and then execute suitable physiological and social behaviours in response. In this review, we provide a comprehensive census of the stimuli and receptors that are linked to modulation of intracellular c-di-GMP. Emerging evidence indicates that c-di-GMP networks sense light, surfaces, energy, redox potential, respiratory electron acceptors, temperature, and structurally diverse biotic and abiotic chemicals. Bioinformatic analysis of sensory domains in diguanylate cyclases and c-di-GMP-specific phosphodiesterases as well as the receptor complexes associated with them reveals that these functions are linked to a diverse repertoire of protein domain families. We describe the principles of stimulus perception learned from studying these modular sensory devices, illustrate how they are assembled in varied combinations with output domains, and summarize a system for classifying these sensor proteins based on their complexity. Biological information-processing via c-di-GMP signal transduction is not only fundamental to bacterial survival in dynamic environments, but also is being used to engineer gene expression circuitry and synthetic proteins with à la carte biochemical functionalities.

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