Cloning and Expression of the Listeria monocytogenes Scott A ptsH and ptsI Genes, Coding for HPr and Enzyme I, Respectively, of the Phosphotransferase System

ABSTRACT The phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) utilizes high-energy phosphate present in PEP to drive the uptake of several different carbohydrates in bacteria. In order to examine the role of the PTS in the physiology of Listeria monocytogenes, we identified the ptsH andptsI genes encoding the HPr and enzyme I proteins, respectively, of the PTS. Nucleotide sequence analysis indicated that the predicted proteins are nearly 70% similar to HPr and enzyme I proteins from other organisms. Purified L. monocytogenesHPr overexpressed in Escherichia coli was also capable of complementing an HPr defect in heterologous extracts ofStaphylococcus aureus ptsH mutants. Additional studies of the transcriptional organization and control indicated that theptsH and ptsI genes are organized into a transcription unit that is under the control of a consensus-like promoter and that expression of these genes is mediated by glucose availability and pH or by by-products of glucose metabolism.

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A Novel Role for Enzyme I of the Vibrio cholerae Phosphoenolpyruvate Phosphotransferase System in Regulation of Growth in a Biofilm

Journal of Bacteriology ◽

10.1128/jb.01410-07 ◽

2007 ◽

Vol 190 (1) ◽

pp. 311-320 ◽

Cited By ~ 56

Author(s):

Laetitia Houot ◽

Paula I. Watnick

Keyword(s):

Vibrio Cholerae ◽

Sugar Transport ◽

Phosphotransferase System ◽

Potent Inducer ◽

Inducer Exclusion ◽

Genes Encoding ◽

Cell Components ◽

Specific Regulation ◽

Enzyme I

ABSTRACT Glucose is a universal energy source and a potent inducer of surface colonization for many microbial species. Highly efficient sugar assimilation pathways ensure successful competition for this preferred carbon source. One such pathway is the phosphoenolpyruvate phosphotransferase system (PTS), a multicomponent sugar transport system that phosphorylates the sugar as it enters the cell. Components required for transport of glucose through the PTS include enzyme I, histidine protein, enzyme IIAGlc, and enzyme IIBCGlc. In Escherichia coli, components of the PTS fulfill many regulatory roles, including regulation of nutrient scavenging and catabolism, chemotaxis, glycogen utilization, catabolite repression, and inducer exclusion. We previously observed that genes encoding the components of the Vibrio cholerae PTS were coregulated with the vps genes, which are required for synthesis of the biofilm matrix exopolysaccharide. In this work, we identify the PTS components required for transport of glucose and investigate the role of each of these components in regulation of biofilm formation. Our results establish a novel role for the phosphorylated form of enzyme I in specific regulation of biofilm-associated growth. As the PTS is highly conserved among bacteria, the enzyme I regulatory pathway may be relevant to a number of biofilm-based infections.

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The role of enzyme I in the unmasking of an essential thiol of the membrane-bound enzyme II of the phosphoenolpyruvate-glucose phosphotransferase system of Escherichia coli

Biochimica et Biophysica Acta (BBA) - Biomembranes ◽

10.1016/0005-2736(77)90183-3 ◽

1977 ◽

Vol 469 (2) ◽

pp. 211-215 ◽

Cited By ~ 10

Author(s):

Rosine Haguenauer-Tsapis ◽

Adam Kepes

Keyword(s):

Escherichia Coli ◽

Phosphotransferase System ◽

Membrane Bound ◽

Enzyme I

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Studies of the Listeria monocytogenes Cellobiose Transport Components and Their Impact on Virulence Gene Repression

Journal of Molecular Microbiology and Biotechnology ◽

10.1159/000500090 ◽

2019 ◽

Vol 29 (1-6) ◽

pp. 10-26 ◽

Cited By ~ 2

Author(s):

Thanh Nguyen Cao ◽

Philippe Joyet ◽

Francine Moussan Désirée Aké ◽

Eliane Milohanic ◽

Josef Deutscher

Keyword(s):

Listeria Monocytogenes ◽

Double Mutant ◽

Virulence Gene ◽

Gene Repression ◽

Phosphotransferase System ◽

Bacteria Transport ◽

Virulence Gene Expression ◽

Transcription Regulator ◽

Related Phenotype

Background: Many bacteria transport cellobiose via a phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS). In Listeria monocytogenes, two pairs of soluble PTS components (EIIACel1/EIIBCel1 and EIIACel2/EIIBCel2) and the permease EIICCel1 were suggested to contribute to cellobiose uptake. Interestingly, utilization of several carbohydrates, including cellobiose, strongly represses virulence gene expression by inhibiting PrfA, the virulence gene activator. Results: The LevR-like transcription regulator CelR activates expression of the cellobiose-induced PTS operons celB1-celC1-celA1, celB2-celA2, and the EIIC-encoding monocistronic celC2. Phosphorylation by P∼His-HPr at His550 activates CelR, whereas phosphorylation by P∼EIIBCel1 or P∼EIIBCel2 at His823 inhibits it. Replacement of His823 with Ala or deletion of both celA or celB genes caused constitutive CelR regulon expression. Mutants lacking EIICCel1, CelR or both EIIACel exhibitedslow cellobiose consumption. Deletion of celC1 or celR prevented virulence gene repression by the disaccharide, but not by glucose and fructose. Surprisingly, deletion of both celA genes caused virulence gene repression even during growth on non-repressing carbohydrates. No cellobiose-related phenotype was found for the celC2 mutant. Conclusion: The two EIIA/BCel pairs are similarly efficient as phosphoryl donors in EIICCel1-catalyzed cellobiose transport and CelR regulation. The permanent virulence gene repression in the celA double mutant further supports a role of PTSCel components in PrfA regulation.

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PTS-Mediated Regulation of the Transcription Activator MtlR from Different Species: Surprising Differences despite Strong Sequence Conservation

Journal of Molecular Microbiology and Biotechnology ◽

10.1159/000369619 ◽

2015 ◽

Vol 25 (2-3) ◽

pp. 94-105 ◽

Cited By ~ 8

Author(s):

Philippe Joyet ◽

Meriem Derkaoui ◽

Houda Bouraoui ◽

Josef Deutscher

Keyword(s):

Bacillus Subtilis ◽

Lactobacillus Casei ◽

Sequence Conservation ◽

Geobacillus Stearothermophilus ◽

Transcription Activator ◽

Phosphotransferase System ◽

Transcription Regulator ◽

Regulatory Domains ◽

Genes Encoding ◽

Enzyme I

The hexitol <smlcap>D</smlcap>-mannitol is transported by many bacteria via a phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS). In most Firmicutes, the transcription activator MtlR controls the expression of the genes encoding the <smlcap>D</smlcap>-mannitol-specific PTS components and <smlcap>D</smlcap>-mannitol-1-P dehydrogenase. MtlR contains an N-terminal helix-turn-helix motif followed by an Mga-like domain, two PTS regulation domains (PRDs), an EIIBGat- and an EIIAMtl-like domain. The four regulatory domains are the target of phosphorylation by PTS components. Despite strong sequence conservation, the mechanisms controlling the activity of MtlR from Lactobacillus casei, Bacillus subtilis and Geobacillus stearothermophilus are quite different. Owing to the presence of a tyrosine in place of the second conserved histidine (His) in PRD2, L. casei MtlR is not phosphorylated by Enzyme I (EI) and HPr. When the corresponding His in PRD2 of MtlR from B. subtilis and G. stearothermophilus was replaced with alanine, the transcription regulator was no longer phosphorylated and remained inactive. Surprisingly, L. casei MtlR functions without phosphorylation in PRD2 because in a ptsI (EI) mutant MtlR is constitutively active. EI inactivation prevents not only phosphorylation of HPr, but also of the PTSMtl components, which inactivate MtlR by phosphorylating its EIIBGat- or EIIAMtl-like domain. This explains the constitutive phenotype of the ptsI mutant. The absence of EIIBMtl-mediated phosphorylation leads to induction of the L. caseimtl operon. This mechanism resembles mtlARFD induction in G. stearothermophilus, but differs from EIIAMtl-mediated induction in B. subtilis. In contrast to B. subtilis MtlR, L. casei MtlR activation does not require sequestration to the membrane via the unphosphorylated EIIBMtl domain.

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Expression of mptC of Listeria monocytogenes induces sensitivity to class IIa bacteriocins in Lactococcus lactis

Microbiology ◽

10.1099/mic.0.27002-0 ◽

2004 ◽

Vol 150 (8) ◽

pp. 2663-2668 ◽

Cited By ~ 75

Author(s):

Manilduth Ramnath ◽

Safia Arous ◽

Anne Gravesen ◽

John W. Hastings ◽

Yann Héchard

Keyword(s):

Lactic Acid ◽

Listeria Monocytogenes ◽

Lactic Acid Bacteria ◽

Lactococcus Lactis ◽

Inhibitory Activity ◽

Bacteriocin Activity ◽

Phosphotransferase System ◽

Enterocin A ◽

Leucocin A

Sensitivity to class IIa bacteriocins from lactic acid bacteria was recently associated with the mannose phosphotransferase system (PTS) permease, , in Listeria monocytogenes. To assess the involvement of this protein complex in class IIa bacteriocin activity, the mptACD operon, encoding , was heterologously expressed in an insensitive species, namely Lactococcus lactis, using the NICE double plasmid system. Upon induction of the cloned operon, the recombinant Lc. lactis became sensitive to leucocin A. Pediocin PA-1 and enterocin A also showed inhibitory activity against Lc. lactis cultures expressing mptACD. Furthermore, the role of the three genes of the mptACD operon was investigated. Derivative plasmids containing various combinations of these three genes were made from the parental mptACD plasmid by divergent PCR. The results showed that expression of mptC alone is sufficient to confer sensitivity to class IIa bacteriocins in Lc. lactis.

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CD44-Regulated Intracellular Proliferation of Listeria monocytogenes

Infection and Immunity ◽

10.1128/iai.71.7.4102-4111.2003 ◽

2003 ◽

Vol 71 (7) ◽

pp. 4102-4111 ◽

Cited By ~ 9

Author(s):

Emma Eriksson ◽

Lone Dons ◽

Antonio Gigliotti Rothfuchs ◽

Paraskevi Heldin ◽

Hans Wigzell ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Intracellular Growth ◽

Murine Bone Marrow ◽

Bacterial Proliferation ◽

Serovar Typhimurium ◽

Inflammatory Processes ◽

And Control ◽

Intracellular Proliferation ◽

Hyaluronan Binding

ABSTRACT CD44 has been implicated in immune and inflammatory processes. We have analyzed the role of CD44 in the outcome of Listeria monocytogenes infection in murine bone marrow-derived macrophages (BMM). Surprisingly, a dramatically decreased intracellular survival of L. monocytogenes was observed in CD44−/− BMM. CD44−/− heart or lung fibroblast cultures also showed reduced bacterial levels. Moreover, livers from CD44−/−-infected mice showed diminished levels of L. monocytogenes. In contrast, intracellular growth of Salmonella enterica serovar Typhimurium was the same in CD44−/− and control BMM. The CD44-mediated increased bacterial proliferation was not linked to altered BMM differentiation or to secretion of soluble factors. CD44 did not mediate listerial uptake, and it played no role in bacterial escape from the primary phagosome or formation of actin tails. Furthermore, CD44-enhanced listerial proliferation occurred in the absence of intracellular bacterial spreading. Interestingly, coincubation of BMM with hyaluronidase or anti-CD44 antibodies that selectively inhibit hyaluronan binding increased intracellular listerial proliferation. Treatment of cells with hyaluronan, in contrast, diminished listerial growth and induced proinflammatory transcript levels. We suggest that L. monocytogenes takes advantage of the CD44-mediated signaling to proliferate intracellularly, although binding of CD44 to certain ligands will inhibit such response.

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Distribution and Functions of Phosphotransferase System Genes in the Genome of the Lactic Acid Bacterium Oenococcus oeni

Applied and Environmental Microbiology ◽

10.1128/aem.00380-13 ◽

2013 ◽

Vol 79 (11) ◽

pp. 3371-3379 ◽

Cited By ~ 13

Author(s):

Zohra Jamal ◽

Cécile Miot-Sertier ◽

François Thibau ◽

Lucie Dutilh ◽

Aline Lonvaud-Funel ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacterium ◽

Oenococcus Oeni ◽

Phosphotransferase System ◽

Content Type ◽

Low Concentrations ◽

Perfect Adaptation ◽

Form Part ◽

Genes Encoding ◽

Enzyme I

ABSTRACTOenococcus oeni, the lactic acid bacterium primarily responsible for malolactic fermentation in wine, is able to grow on a large variety of carbohydrates, but the pathways by which substrates are transported and phosphorylated in this species have been poorly studied. We show that the genes encoding the general phosphotransferase proteins, enzyme I (EI) and histidine protein (HPr), as well as 21 permease genes (3 isolated ones and 18 clustered into 6 distinct loci), are highly conserved among the strains studied and may form part of theO. oenicore genome. Additional permease genes differentiate the strains and may have been acquired or lost by horizontal gene transfer events. The coreptsgenes are expressed, and permease gene expression is modulated by the nature of the bacterial growth substrate. DecryptifiedO. oenicells are able to phosphorylate glucose, cellobiose, trehalose, and mannose at the expense of phosphoenolpyruvate. These substrates are present at low concentrations in wine at the end of alcoholic fermentation. The phosphotransferase system (PTS) may contribute to the perfect adaptation ofO. oenito its singular ecological niche.

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Regulation of ABCA1 and ABCG1 gene expression in the intraabdominal adipose tissue

Biomeditsinskaya Khimiya ◽

10.18097/pbmc20166203283 ◽

2016 ◽

Vol 62 (3) ◽

pp. 283-289 ◽

Cited By ~ 2

Author(s):

V.V. Miroshnikova ◽

A.A. Panteleeva ◽

E.A. Bazhenova ◽

E.P. Demina ◽

T.S. Usenko ◽

...

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Cholesterol Metabolism ◽

Nuclear Factor ◽

Specific Expression ◽

Protein Levels ◽

Expression Of Genes ◽

Genes Encoding ◽

And Control

Tissue specific expression of genes encoding cholesterol transporters ABCA1 and ABCG1 as well as genes encoding the most important transcriptional regulators of adipogenesis – LXRa, LXRb, PPARg and RORa has been investigated in intraabdominal adipose tissue (IAT) samples.A direct correlation between the content of ABCA1 and ABCG1 proteins with RORa protein level (r=0.480, p<0.05; r=0.435, p<0.05, respectively) suggests the role of the transcription factor RORa in the regulation of IAT ABCA1 and ABCG1 protein levels. ABCA1 and ABCG1 gene expression positively correlated with obesity indicators such as body mass index (BMI) (r=0.522, p=0.004; r=0.594, p=0.001, respectively) and waist circumference (r=0.403, p=0.033; r=0.474, p=0.013, respectively). The development of obesity is associated with decreased IAT levels of RORa and LXRb mRNA (p=0.016 and p=0.002, respectively). These data suggest that the nuclear factor RORa can play a significant role in the regulation of cholesterol metabolism and control IAT expression of ABCA1 and ABCG1, while the level of IAT LXRb gene expression may be an important factor associated with the development of obesity.

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Glycerol Metabolism and PrfA Activity in Listeria monocytogenes

Journal of Bacteriology ◽

10.1128/jb.00259-08 ◽

2008 ◽

Vol 190 (15) ◽

pp. 5412-5430 ◽

Cited By ~ 79

Author(s):

Biju Joseph ◽

Sonja Mertins ◽

Regina Stoll ◽

Jennifer Schär ◽

Kanasinakatte Rudrappa Umesha ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Glycerol Metabolism ◽

Phosphotransferase System ◽

Genes Encoding ◽

N Metabolism ◽

Branched Chain ◽

Glycerol Uptake ◽

High Level ◽

Uptake And Metabolism ◽

Logarithmic Growth

ABSTRACT Listeria monocytogenes is able to efficiently utilize glycerol as a carbon source. In a defined minimal medium, the growth rate (during balanced growth) in the presence of glycerol is similar to that in the presence of glucose or cellobiose. Comparative transcriptome analyses of L. monocytogenes showed high-level transcriptional upregulation of the genes known to be involved in glycerol uptake and metabolism (glpFK and glpD) in the presence of glycerol (compared to that in the presence of glucose and/or cellobiose). Levels of expression of the genes encoding a second putative glycerol uptake facilitator (GlpF2) and a second putative glycerol kinase (GlpK2) were less enhanced under these conditions. GlpK1 but not GlpK2 was essential for glycerol catabolism in L. monocytogenes under extracellular conditions, while the loss of GlpK1 affected replication in Caco-2 cells less than did the loss of GlpK2 and GlpD. Additional genes whose transcription levels were higher in the presence of glycerol than in the presence of glucose and cellobiose included those for two dihydroxyacetone (Dha) kinases and many genes that are under carbon catabolite repression control. Transcriptional downregulation in the presence of glycerol (compared to those in the presence glucose and cellobiose) was observed for several genes and operons that are positively regulated by glucose, including genes involved in glycolysis, N metabolism, and the biosynthesis of branched-chain amino acids. The highest level of transcriptional upregulation was observed for all PrfA-dependent genes during early and late logarithmic growth in glycerol. Under these conditions, a low level of HPr-Ser-P and a high level of HPr-His-P were present in the cells, suggesting that all enzyme IIA (EIIA) (or EIIB) components of the phosphotransferase system (PTS) permeases expressed will be phosphorylated. These and other data suggest that the phosphorylation state of PTS permeases correlates with PrfA activity.

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The role of phosphotransferase-mediated syntheses of fructose 1-phosphate and fructose 6-phosphate in the growth of Escherichia coli on fructose

Proceedings of the Royal Society of London Series B Biological Sciences ◽

10.1098/rspb.1974.0065 ◽

1974 ◽

Vol 187 (1087) ◽

pp. 105-119 ◽

Cited By ~ 29

Keyword(s):

Escherichia Coli ◽

Kinase Activity ◽

Phosphotransferase System ◽

High Concentrations ◽

Enzyme I ◽

Uptake And Metabolism

Mutants of Escherichia coli impaired in fructose 1-phosphate kinase activity (Fpk) accumulate fructose 1-phosphate from fructose, which arrests their growth. Phenotypic revertants to fructose tolerance have lost either the histidine-containing protein (HPr) or enzyme I of the phosphoenolpyruvate phosphotransferase system ( ctr ), and consequently utilize neither fructose nor glucose, or an enzyme II specific for the uptake of fructose and its concomitant phosphorylation to fructose 1-phosphate (PtsF). However, PtsF — -mutants can still grow on high concentrations ( > 2 mM) of fructose, and take up this sugar via a low-affinity enzyme II designated PtsX that effects its uptake and phosphorylation to fructose 6-phosphate. Mutants of the Hfr-strain KL16 were isolated that lacked PtsF, or PtsF and PtsX, activities. The consequence to, and rôle of these functions in, the uptake and metabolism of fructose are described.

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