The major PEP-phosphotransferase systems (PTSs) for glucose, mannose and cellobiose of Listeria monocytogenes, and their significance for extra- and intracellular growth

Stoll, Regina; Goebel, Werner

doi:10.1099/mic.0.034934-0

Cited by 83 publications

(107 citation statements)

References 34 publications

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“…S1F in the supplemental material). This gene encodes the transcriptional regulator LacR, which has been suggested to suppress virulence in L. monocytogenes in response to cellobiose (54,55) in addition to regulating the phosphotransferase system (56). Plaque formation was unaffected by in-frame deletion of the lmo1721 gene alone or in combination with a secA2 deletion (Fig.…”

Section: Resultsmentioning

confidence: 93%

A prl Mutation in SecY Suppresses Secretion and Virulence Defects of Listeria monocytogenes secA2 Mutants

2015

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The bulk of bacterial protein secretion occurs through the conserved SecY translocation channel that is powered by SecA-dependent ATP hydrolysis. Many Gram-positive bacteria, including the human pathogen Listeria monocytogenes, possess an additional nonessential specialized ATPase, SecA2. SecA2-dependent secretion is required for normal cell morphology and virulence in L. monocytogenes; however, the mechanism of export via this pathway is poorly understood. L. monocytogenes secA2 mutants form rough colonies, have septation defects, are impaired for swarming motility, and form small plaques in tissue culture cells. In this study, 70 spontaneous mutants were isolated that restored swarming motility to L. monocytogenes secA2 mutants. Most of the mutants had smooth colony morphology and septated normally, but all were lysozyme sensitive. Five representative mutants were subjected to whole-genome sequencing. Four of the five had mutations in proteins encoded by the lmo2769 operon that conferred lysozyme sensitivity and increased swarming but did not rescue virulence defects. A point mutation in secY was identified that conferred smooth colony morphology to secA2 mutants, restored wild-type plaque formation, and increased virulence in mice. This secY mutation resembled a prl suppressor known to expand the repertoire of proteins secreted through the SecY translocation complex. Accordingly, the ⌬secA2prlA1 mutant showed wild-type secretion levels of P60, an established SecA2-dependent secreted autolysin. Although the prl mutation largely suppressed almost all of the measurable SecA2-dependent traits, the ⌬secA2prlA1 mutant was still less virulent in vivo than the wild-type strain, suggesting that SecA2 function was still required for pathogenesis.T he essential general secretory (Sec) pathway is responsible for exporting the majority of secreted proteins across the bacterial cell membrane (1-3). Much of what we know about this pathway was discovered in Escherichia coli by using genetic screens to identify loss-of-function mutations in components of the Sec system. This led to the identification of components of the SecYEG complex that forms the translocation channel and the SecA ATPase that binds to signal sequences to drive the export of precursor proteins across the channel (3-9). In contrast to loss-of-function sec mutations, prl alleles are gain-of-function mutations, which expand the repertoire of substrates being exported across the SecYEG channel (6-8, 10, 11). The most dominant prl variants are in secY, known as prlA mutations, which allow for the secretion of proteins with altered signal sequence (7, 9, 11) without significantly altering the secretion of other proteins (6,12).In addition to the canonical SecA, an accessory cytosolic ATPase, SecA2, has been identified in Mycobacterium and a subset of other Gram-positive bacteria. Unlike SecA, SecA2 is not essential for cell viability but is required for virulence in some pathogenic bacteria (13-15). SecA1 and SecA2 are homologous but are not interchangeable (16)...

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Section: Resultsmentioning

confidence: 93%

A prl Mutation in SecY Suppresses Secretion and Virulence Defects of Listeria monocytogenes secA2 Mutants

2015

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“…This protein controls the expression of the manLMNO operon, which encodes the major glucose transporter of this human pathogen (78,150). However, ManR activity is regulated not by the components of the mannose-type PTS ManLMN but by the proteins of another mannose-type PTS, MpoABCD, a low-affinity glucose/mannose-specific PTS that functions mainly as glucose sensor (78 PTS components also play a role in biofilm formation, as has been shown for EI and HPr of V. cholerae (151).…”

Section: Fig 5 Pts-catalyzed Glucose Uptake and The Eiiamentioning

confidence: 99%

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Deutscher

Aké

Derkaoui

et al. 2014

Microbiol Mol Biol Rev

341

374

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SUMMARY The bacterial phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) carries out both catalytic and regulatory functions. It catalyzes the transport and phosphorylation of a variety of sugars and sugar derivatives but also carries out numerous regulatory functions related to carbon, nitrogen, and phosphate metabolism, to chemotaxis, to potassium transport, and to the virulence of certain pathogens. For these different regulatory processes, the signal is provided by the phosphorylation state of the PTS components, which varies according to the availability of PTS substrates and the metabolic state of the cell. PEP acts as phosphoryl donor for enzyme I (EI), which, together with HPr and one of several EIIA and EIIB pairs, forms a phosphorylation cascade which allows phosphorylation of the cognate carbohydrate bound to the membrane-spanning EIIC. HPr of firmicutes and numerous proteobacteria is also phosphorylated in an ATP-dependent reaction catalyzed by the bifunctional HPr kinase/phosphorylase. PTS-mediated regulatory mechanisms are based either on direct phosphorylation of the target protein or on phosphorylation-dependent interactions. For regulation by PTS-mediated phosphorylation, the target proteins either acquired a PTS domain by fusing it to their N or C termini or integrated a specific, conserved PTS regulation domain (PRD) or, alternatively, developed their own specific sites for PTS-mediated phosphorylation. Protein-protein interactions can occur with either phosphorylated or unphosphorylated PTS components and can either stimulate or inhibit the function of the target proteins. This large variety of signal transduction mechanisms allows the PTS to regulate numerous proteins and to form a vast regulatory network responding to the phosphorylation state of various PTS components.

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“…When the PTS Man is inactivated, L. monocytogenes still utilizes glucose at ϳ30% of the efficiency of the wild-type strain (26). This remaining activity is due mostly to a second PTS of the mannose class (27) (15,26). In L. monocytogenes, several other PTS and non-PTS transporters are able to take up glucose with low affinity, most notably a PtsG-like permease of the glucose class, which is encoded by lmo0027.…”

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confidence: 99%

“…It has the capacity to switch from a saprophytic to a virulent life-style (13). In order to exploit the numerous carbon sources in soil derived from decaying plants and other organisms, L. monocytogenes possesses a large number of carbohydrate transporters (14), many of which belong to the PTS (15). The utilization of an efficiently metabolized PTS substrate was suggested to indicate to the bacterium its presence in a saprophytic environment, because it represses the expression of most virulence genes (16).…”

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confidence: 99%

Interaction with Enzyme IIB^Mpo(EIIB^Mpo) and Phosphorylation by Phosphorylated EIIB^MpoExert Antagonistic Effects on the Transcriptional Activator ManR of Listeria monocytogenes

et al. 2015

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Listeriae take up glucose and mannose predominantly through a mannose class phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS Man ), whose three components are encoded by the manLMN genes. Mpo and PϳPRD2 of ManR, which together lead to the induction of the manLMN operon. Complementation of a ⌬manR mutant with various manR alleles confirmed the antagonistic effects of PTS-catalyzed phosphorylation at the two different histidine residues of ManR. Deletion of manR prevented not only the expression of the manLMN operon but also glucose-mediated repression of virulence gene expression; however, repression by other carbohydrates was unaffected. Interestingly, the expression of manLMN in Listeria innocua was reported to require not only ManR but also the Crp-like transcription activator Lin0142. Unlike Lin0142, the L. monocytogenes homologue, Lmo0095, is not required for manLMN expression; its absence rather stimulates man expression. IMPORTANCEListeria monocytogenes is a human pathogen causing the foodborne disease listeriosis. The expression of most virulence genes is controlled by the transcription activator PrfA. Its activity is strongly repressed by carbohydrates, including glucose, which is transported into L. monocytogenes mainly via a mannose/glucose-specific phosphotransferase system (PTS Man ). Expression of the man operon is regulated by the transcription activator ManR, the activity of which is controlled by a second, low-efficiency PTS of the mannose family, which functions as glucose sensor. Here we demonstrate that the EIIB Mpo component plays a dual role in ManR regulation: it inactivates ManR by phosphorylating its His871 residue and stimulates ManR by interacting with its two C-terminal domains. Glucose is recognized as a preferred carbon source for numerous bacteria. When multiple carbohydrates are present in a growth medium, glucose represses the synthesis of the enzymes required for the utilization of other carbon sources and is therefore the first to be transported and metabolized. This phenomenon is known as carbon catabolite repression. Glucose is taken up by bacterial cells either by ion-driven permeases, such as GlcP (1) and GlcU (2), or via the phosphoenolpyruvate (PEP):glycose phosphotransferase system (PTS), with the latter being the most common bacterial glucose transporter. The PTS is composed of four proteins or protein domains, which form a phosphorylation cascade, and one or two membrane-spanning proteins (3). The phosphorylation cascade starts with the PEP-requiring autophosphorylation of a histidyl residue in enzyme I (EI) (4). Phosphorylated EI (PϳEI) then transfers the phosphoryl group to histidine 15 in HPr, the second general PTS protein (Fig. 1). In the next step, PϳHis-HPr phosphorylates a histidyl residue in a carbohydrate-specific EIIA component; bacteria usually contain several EIIA components. PϳEIIA subsequently phosphorylates a cysteyl or histidyl residue in the EIIB component of the same

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The major PEP-phosphotransferase systems (PTSs) for glucose, mannose and cellobiose of Listeria monocytogenes, and their significance for extra- and intracellular growth

Cited by 83 publications

References 34 publications

A prl Mutation in SecY Suppresses Secretion and Virulence Defects of Listeria monocytogenes secA2 Mutants

A prl Mutation in SecY Suppresses Secretion and Virulence Defects of Listeria monocytogenes secA2 Mutants

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Interaction with Enzyme IIB^Mpo(EIIB^Mpo) and Phosphorylation by Phosphorylated EIIB^MpoExert Antagonistic Effects on the Transcriptional Activator ManR of Listeria monocytogenes

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The major PEP-phosphotransferase systems (PTSs) for glucose, mannose and cellobiose of Listeria monocytogenes, and their significance for extra- and intracellular growth

Cited by 83 publications

References 34 publications

A prl Mutation in SecY Suppresses Secretion and Virulence Defects of Listeria monocytogenes secA2 Mutants

A prl Mutation in SecY Suppresses Secretion and Virulence Defects of Listeria monocytogenes secA2 Mutants

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Interaction with Enzyme IIBMpo(EIIBMpo) and Phosphorylation by Phosphorylated EIIBMpoExert Antagonistic Effects on the Transcriptional Activator ManR of Listeria monocytogenes

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Interaction with Enzyme IIB^Mpo(EIIB^Mpo) and Phosphorylation by Phosphorylated EIIB^MpoExert Antagonistic Effects on the Transcriptional Activator ManR of Listeria monocytogenes