The cell wall precursor lipid II acts as a molecular signal for the Ser/Thr kinase PknB of Staphylococcus aureus

Hardt, Patrick; Engels, Ina; Rausch, Marvin D.; Gajdiss, Mike; Ulm, Hannah; Saß, Peter; Ohlsen, Knut; Sahl, Hans‐Georg; Bierbaum, Gabriele; Schneider, Tanja; Grein, Fabian

doi:10.1016/j.ijmm.2016.12.001

Cited by 76 publications

(94 citation statements)

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“…8). It has been demonstrated previously that the kinase activity of PrkA homologues was activated by muropeptides (17, 49) or the PG precursor lipid II (18). Muropeptides were released from the cell wall during normal PG turnover, and their release was intensified when PG hydrolysis prevailed over PG biosynthesis (10, 50), whereas blocking PG chain elongation by moenomycin treatment caused the accumulation of lipid-linked PG precursors (51).…”

Section: Discussionmentioning

confidence: 98%

“…These membrane-integral enzymes comprise a cytoplasmic kinase domain linked to several extracellular PASTA domains by a single transmembrane region (15). The kinase activity is stimulated by free muropeptides and lipid II (that accumulate during damage and turnover of PG) on interaction with the PASTA domains (17, 18). PknB, a representative of this protein family from Mycobacterium tuberculosis , phosphorylates GlmU, a bifunctional uridyltransferase/acetyltransferase important for synthesis of UDP-Glc N Ac, and in so doing reduces GlmU activity (19).…”

Section: Introductionmentioning

confidence: 99%

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PrkA controls peptidoglycan biosynthesis through the essential phosphorylation of ReoM

Wamp

Rutter

Rismondo

et al. 2019

Preprint

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30Peptidoglycan (PG) is the main component of bacterial cell walls and the target for many 31 antibiotics. PG biosynthesis is tightly coordinated with cell wall growth and turnover and many 32 of these control activities depend upon PASTA-domain containing eukaryotic-like 33 serine/threonine protein kinases (PASTA-eSTK) that sense PG fragments. However, only a few 34 PG biosynthetic enzymes are actually direct kinase substrates. Here, we identify the conserved 35 ReoM protein as a novel PASTA-eSTK substrate in the Gram-positive pathogen Listeria 36 monocytogenes. Our data show that the phosphorylation of ReoM is essential as it controls 37 ClpCP-dependent proteolytic degradation of the essential enzyme MurA, which catalyses the first 38 committed step in PG biosynthesis. We also identify ReoY as a second novel factor required for 39 degradation of ClpCP substrates. Collectively, our data imply that the first committed step of PG 40 biosynthesis is activated through control of ClpCP protease activity in response to signals of PG 41 homeostasis imbalance.42 43 44 45The cell wall of Gram-positive bacteria is a complicated three-dimensional structure that engulfs 46 the cell as a closed sacculus. The main component of bacterial cell walls is peptidoglycan (PG), a 47 network of glycan strands crosslinked together by short peptides (1). PG biosynthesis starts with 48 the conversion of UDP-GlcNAc into lipid II, a disaccharide pentapeptide that is ligated to a 49 100 positive bacteria, which explains how PG biosynthesis is adjusted in these bacteria to meet PG 101 production and repair needs. 102 103 6 RESULTS 104 105Novel gpsB suppressor mutations in the lmo1503 (reoM) and lmo1921 (reoY) genes 106 A L. monocytogenes ΔgpsB mutant is unable to replicate at 42°C but forms suppressors 107 correcting this defect with high frequency (28). In a preliminary selection of gpsB suppressors, 108 the clpC and murZ genes, important for the stability of the UDP-N-acetylglucosamine 1-109 carboxyvinyltransferase MurA, were found to be mutated (28). We have characterized three more 110 shg (suppression of heat sensitive growth) suppressor mutants (shg8, shg10 and shg12) isolated 111 from a ΔgpsB mutant incubated on a BHI agar plate at 42°C. These three shg strains grew as fast 112 as the wild type when cultivated at 37°C or 42°C, whereas the parental ΔgpsB mutant grew at a 113 reduced rate at 37°C and did not grow at 42°C (Fig. 1A-B), as shown previously (32). 114Illumina sequencing of the shg8, shg10 and shg12 genomes identified one SNP in each strain that 115 was not found in the parental ΔgpsB strain. Strain shg8 carried a mutation in the uncharacterized 116 lmo1921 gene (herein named reoY, see below) that exchanged H87 into tyrosine; the same gene 117 was affected by the introduction of a premature stop codon after the 73 rd codon of reoY to yield 118 strain shg10. Strain shg12 carried a mutation in the ribosomal binding site (RBS) of the lmo1503 119 gene (renamed reoM), encoding another protein of unknown function and conta...

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

PrkA controls peptidoglycan biosynthesis through the essential phosphorylation of ReoM

Wamp

Rutter

Rismondo

et al. 2019

Preprint

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“…The latter two strains were included to differentiate the site of phosphorylation. PknB has been shown to phosphorylate WalR at residue T101 37 while in B. subtilis, the deletion of Stpl led to increased WalR phosphorylation at the site equivalent to WalR T101 38 . Here, we observed that D53 phosphorylation on WalR was absent in the WalR D 53 FLAG (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…These include a second site of phosphorylation, residue T101 on WaIR, by the serine threonine kinase known as Stkl or PknB 37 , and the interaction of SpdC (previously called lysostaphin resistance factor A – lyrA) 44 with WalK, which negatively regulate genes under the control of WaIR 45 . Both PknB and SpdC are localised to the division septum similar to WalK 31,37 , highlighting the complexity of this regulatory axis. The establishment of phos-tag acrylamide to analyse the phosphorylation status of WalR in vivo is a powerful tool to investigate the regulation of this essential system (Fig.…”

Section: Discussionmentioning

confidence: 99%

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

Monk

Shaikh

Begg

et al. 2018

Preprint

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WalKR (YycFG) is the only essential two-component regulator in the human pathogen Staphylococcus aureus. WalKR regulates peptidoglycan synthesis, but this function alone appears not to explain its essentiality. To understand WalKR function we investigated a suppressor mutant that arose when WalKR activity was impaired; a single histidine to tryptophan substitution (H271Y) in the cytoplasmic Per-Arnt-Sim (PASCYT) domain of the histidine kinase WalK. Introduction of the WalKH271Y mutation into wild-type S. aureus activated the WalKR regulon. Structural analyses of the WalK PASCYT domain revealed a hitherto unknown metal binding site, in which a zinc ion (Zn2+) was tetrahedrally-coordinated by four amino acid residues including H271. The WallkH271Y mutation abrogated metal binding, increasing WalK kinase activity and WalR phosphorylation. Thus, Zn2+-binding negatively regulates WalKR activity. Identification of a metal ligand sensed by the WalKR system substantially expands our understanding of this critical S. aureus regulon.

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“…The S. aureus Stk1 Hanks kinase is also involved in the regulation of cell wall metabolism and division [32]. The cell wall precursor molecule Lipid II has been reported as the activating signal for Stk1, and binding to lipidII variants induces Stk1 autophosphorylation in vitro [46]. In vivo, Stk1 localizes to the septum following recruitment of the contractile Z-ring forming protein FtsZ.…”

Section: Pathogen Cell Division and Cell Wall Biosynthesismentioning

confidence: 99%

Importance of protein Ser/Thr/Tyr phosphorylation for bacterial pathogenesis

et al. 2020

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Protein phosphorylation regulates a large variety of biological processes in all living cells. In pathogenic bacteria, the study of serine, threonine, and tyrosine (Ser/Thr/Tyr) phosphorylation has shed light on the course of infectious diseases, from adherence to host cells to pathogen virulence, replication, and persistence. Mass spectrometry (MS)-based phosphoproteomics has provided global maps of Ser/Thr/Tyr phosphosites in bacterial pathogens. Despite recent developments, a quantitative and dynamic view of phosphorylation events that occur during bacterial pathogenesis is currently lacking. Temporal, spatial, and subpopulation resolution of phosphorylation data is required to identify key regulatory nodes underlying bacterial pathogenesis. Herein, we discuss how technological improvements in sample handling, MS instrumentation, data processing, and machine learning should improve bacterial phosphoproteomic datasets and the information extracted from them. Such information is expected to significantly extend the current knowledge of Ser/ Thr/Tyr phosphorylation in pathogenic bacteria and should ultimately contribute to the design of novel strategies to combat bacterial infections.

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The cell wall precursor lipid II acts as a molecular signal for the Ser/Thr kinase PknB of Staphylococcus aureus

Cited by 76 publications

References 61 publications

PrkA controls peptidoglycan biosynthesis through the essential phosphorylation of ReoM

PrkA controls peptidoglycan biosynthesis through the essential phosphorylation of ReoM

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

Importance of protein Ser/Thr/Tyr phosphorylation for bacterial pathogenesis

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