Zinc-binding to the cytoplasmic PAS domain regulates the essential WalK histidine kinase of Staphylococcus aureus

Monk, Ian R.; Shaikh, Nausad H.; Begg, Stephanie L.; Gajdiss, Mike; Sharkey, Liam; Lee, Jean; Pidot, Sacha J.; Seemann, Torsten; Kuiper, Michael J.; Winnen, Brit; Hvorup, Rikki N.; Collins, Brett M.; Bierbaum, Gabriele; Udagedara, Saumya R.; Morey, Jacqueline R.; Pulyani, Neha; Howden, Benjamin P; Maher, M.J.; McDevitt, Christopher A.; King, Glenn F.; Stinear, Timothy P.

doi:10.1038/s41467-019-10932-4

Cited by 41 publications

(50 citation statements)

References 68 publications

(87 reference statements)

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“…WalK has an extracytoplasmic and a cytoplasmic PAS domain, both being potential candidates for sensing ligands that control the phosphorylation activity. Crystallization of the cytoplasmic PAS domain revealed a Zinc atom binding site, which upon zinc-binding leads to a conformational change of WalK that inhibits WalRK phosphorylation [9]. Very recently it was shown that so far unidentified peptidoglycan degradation products modify the activity of B. subtilis WalK and might interact with the extracellular PAS domain [20].…”

Section: Discussionmentioning

confidence: 99%

“…Work in B. subtilis indicates that the extracellular PAS domain might provide feedback information about the activity of the autolysins [20]. Zinc binding to the intracellular PAS domain is special for staphylococci and enterococci and inhibition by zinc was mainly observed at the entry into stationary phase [9], indicating that zinc might act to decrease activity of WalK at this time point. The role of the zinc signaling for S. aureus is unclear, but zinc is essential for S. aureus, a co-factor of some autolytic amidases [52] and plays a role in bacterial infection and innate immunity [53].…”

Section: Discussionmentioning

confidence: 99%

“…Reduction of the protein expression was controlled by Western blotting with specific anti- YycH and anti-YycI antibodies ( Figure 2 ). The pIMAY-ZΔ yycHI deletion vector was constructed as described previously [ 9 ]. The plasmid was transformed into S. aureus NRS384 walR -FLAG with allelic exchange conducted as described by Monk et al [ 27 ].…”

Section: Methodsmentioning

confidence: 99%

“…The plasmid was transformed into S. aureus NRS384 walR-FLAG with allelic exchange conducted as described by Monk et al [27]. S. aureus NRS384 is a member of the USA300 community associated epidemic MRSA [9].…”

Section: Creation Of Yych Yyci and Yychi Antisense Knockdown Clonesmentioning

confidence: 99%

“…In addition to its role in cell wall metabolism, WalRK has been reported to be involved in S. aureus host colonization [ 8 ], virulence [ 2 , 4 ], and biofilm formation [ 3 ]. The highest activity of the WalRK TCS occurs at the end of the log phase in B. subtilis [ 5 ] and in S. aureus [ 9 ] and entails autophosphorylation of the membrane bound WalK kinase, followed by phosphotransfer to the cytoplasmic response regulator WalR. Activation by phosphorylation leads to dimerization and binding to specific promoter regions, thereby altering the transcription level of the corresponding genes.…”

Section: Introductionmentioning

confidence: 99%

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YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation

et al. 2020

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Staphylococcus aureus is a facultative pathogen that can encode numerous antibiotic resistance and immune evasion genes and can cause severe infections. Reduced susceptibility to last resort antibiotics such as vancomycin and daptomycin is often associated with mutations in walRK, an essential two-component regulatory system (TCS). This study focuses on the WalK accessory membrane proteins YycH and YycI and their influence on WalRK phosphorylation. Depletion of YycH and YycI by antisense RNA caused an impaired autolysis, indicating a positive regulatory function on WalK as has been previously described. Phosphorylation assays with full-length recombinant proteins in phospholipid liposomes showed that YycH and YycI stimulate WalK activity and that both regulatory proteins are needed for full activation of the WalK kinase. This was validated in vivo through examining the phosphorylation status of WalR using Phos-tag SDS-PAGE with a yycHI deletion mutant exhibiting reduced levels of phosphorylated WalR. In the yycHI knockdown strain, muropeptide composition of the cell wall was not affected, however, the wall teichoic acid content was increased. In conclusion, a direct modulation of WalRK phosphorylation activity by the accessory proteins YycH and YycI is reported both in vitro and in vivo. Taken together, our results show that YycH and YycI are important in the direct regulation of WalRK-dependent cell wall metabolism.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Creation Of Yych Yyci and Yychi Antisense Knockdown Clonesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation

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Labile Coordination Interphase for Regulating Lean Ion Dynamics in Reversible Zn Batteries

Wang,

Zhu,

Vi‐Tang

et al. 2023

Advanced Materials

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The rechargeable Zn battery technology is plagued with limited reversibility on the anode. The issue is particularly exacerbated when using lean electrolytes, which compromises the economic advantages of zinc batteries for large‐scale energy storage. In this paper, we report the development of a zinc‐coordinated interphase that prevents chemical corrosion and provides protection for zinc anodes. The selective binding of Zn2+ ions towards histidine and carboxylate ligands forms a coordination environment that offers high Zn2+ affinity and fast Zn2+ diffusion owing to its thermodynamic stability and kinetic lability. Both experiments and calculations suggest that the interphase mitigates side reactions and regulates the dendrite‐free electrodeposition. Implementing this labile coordination interphase yields enhanced cycling at a high current density of 20 mA cm−2 with high reversibility of dendrite‐free zinc plating/stripping for more than 200 hours. The reversibility was demonstrated in a Zn||LiMn2O4 cell, which shows an energy density of 74.7 mWh g−1 and a Coulombic efficiency of 99.7% after 500 cycles. In addition, we demonstrate a lean‐electrolyte full cell utilizing electrolyte of only 10 μL mAh−1. This cell operates for an extended cycling life of 100 cycles, five times longer than the pristine Zn anodes. This provides an energy density that is significantly higher than that of commercial aqueous batteries. This work demonstrates a proof‐of‐concept design for utilizing labile coordination interphases on Zn anodes, offering an approach for low‐electrolyte usage and the manufacture of high‐energy‐density batteries.This article is protected by copyright. All rights reserved

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TetR and OmpR family regulators in natural product biosynthesis and resistance

Patil,

Sharma,

Bhaskarwar

et al. 2023

Proteins

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This article provides a comprehensive review and sequence‐structure analysis of transcription regulator (TR) families, TetR and OmpR/PhoB, involved in specialized secondary metabolite (SSM) biosynthesis and resistance. Transcription regulation is a fundamental process, playing a crucial role in orchestrating gene expression to confer a survival advantage in response to frequent environmental stress conditions. This process, coupled with signal sensing, enables bacteria to respond to a diverse range of intra and extracellular signals. Thus, major bacterial signaling systems use a receptor domain to sense chemical stimuli along with an output domain responsible for transcription regulation through DNA‐binding. Sensory and output domains on a single polypeptide chain (one component system, OCS) allow response to stimuli by allostery, that is, DNA‐binding affinity modulation upon signal presence/absence. On the other hand, two component systems (TCSs) allow cross‐talk between the sensory and output domains as they are disjoint and transmit information by phosphorelay to mount a response. In both cases, however, TRs play a central role. Biosynthesis of SSMs, which includes antibiotics, is heavily regulated by TRs as it diverts the cell's resources towards the production of these expendable compounds, which also have clinical applications. These TRs have evolved to relay information across specific signals and target genes, thus providing a rich source of unique mechanisms to explore towards addressing the rapid escalation in antimicrobial resistance (AMR). Here we focus on the TetR and OmpR family TRs, which belong to OCS and TCS, respectively. These TR families are well‐known examples of regulators in secondary metabolism and are ubiquitous across different bacteria, as they also participate in a myriad of cellular processes apart from SSM biosynthesis and resistance. As a result, these families exhibit higher sequence divergence, which is also evident from our bioinformatic analysis of 158 389 and 77 437 sequences from TetR and OmpR family TRs, respectively. The analysis of both sequence and structure allowed us to identify novel motifs in addition to the known motifs responsible for TR function and its structural integrity. Understanding the diverse mechanisms employed by these TRs is essential for unraveling the biosynthesis of SSMs. This can also help exploit their regulatory role in biosynthesis for significant pharmaceutical, agricultural, and industrial applications.

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Zinc-binding to the cytoplasmic PAS domain regulates the essential WalK histidine kinase of Staphylococcus aureus

Cited by 41 publications

References 68 publications

YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation

YycH and YycI Regulate Expression of Staphylococcus aureus Autolysins by Activation of WalRK Phosphorylation

Labile Coordination Interphase for Regulating Lean Ion Dynamics in Reversible Zn Batteries

TetR and OmpR family regulators in natural product biosynthesis and resistance

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