scholarly journals Stress Response and Adaptation of Listeria monocytogenes 08-5923 Exposed to a Sublethal Dose of Carnocyclin A

2014 ◽  
Vol 80 (13) ◽  
pp. 3835-3841 ◽  
Author(s):  
Xiaoji Liu ◽  
Urmila Basu ◽  
Petr Miller ◽  
Lynn M. McMullen
Keyword(s):  
Cell Wall ◽  
Cell Division ◽  
Listeria Monocytogenes ◽  
Stress Responses ◽  
Morphological Changes ◽  
Meat Products ◽  
Sublethal Dose ◽  
Cell Wall Synthesis ◽  
Content Type ◽  
Carnocyclin A

ABSTRACTCarnocyclin A (CCLA) is an antimicrobial peptide produced byCarnobacterium maltaromaticumATCC PTA-5313, which can be used to control the growth ofListeria monocytogenesin ready-to-eat meat products. The aim of this research was to elucidate the cellular responses ofL. monocytogenes08-5923 exposed to a sublethal dose of CCLA. Microarray, quantitative reverse transcription-PCR, tandem mass spectrometry, and electron microscopy were used to investigate the alteration in gene expression, protein production, and morphological changes in cells ofListeriafollowing treatment with CCLA. The genes involved in metabolism (baiE,trn, andpykA), cell wall synthesis (murZanddacB2), and cell division (clpEanddivIVA) were upregulated following a 15-min exposure to CCLA as a result of stress responses. Genes involved in cell division, cell wall synthesis, flagellar synthesis, and metabolism were downregulated after 4 h as a result of adaptation. Analysis of total soluble proteins confirmed the downregulation ofpykAandgndafter 4 h of exposure to CCLA. The absence of flagella was observed inL. monocytogenesfollowing 30 h of exposure to CCLA. A sublethal dose of CCLA induced adaptation inL. monocytogenes08-5923 by inhibition of expression of genes and proteins critical for synthesis of cell wall structures and maintaining metabolic functions. Both the mannose- and cellobiose-specific phosphotransferase systems could be targets for CCLA.

scholarly journals Bacteriophage SP01 Gene Product 56 Inhibits Bacillus subtilis Cell Division by Interacting with FtsL and Disrupting Pbp2B and FtsW Recruitment

2020 ◽  
Vol 203 (2) ◽  
pp. e00463-20
Author(s):  
Amit Bhambhani ◽  
Isabella Iadicicco ◽  
Jules Lee ◽  
Syed Ahmed ◽  
Max Belfatto ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cell Division ◽  
Gene Product ◽  
Fluorescent Protein ◽  
Lytic Bacteriophage ◽  
Cell Wall Synthesis ◽  
Content Type ◽  
Ftsz Ring ◽  
Green Fluorescent

ABSTRACTPrevious work identified gene product 56 (gp56), encoded by the lytic bacteriophage SP01, as being responsible for inhibition of Bacillus subtilis cell division during its infection. Assembly of the essential tubulin-like protein FtsZ into a ring-shaped structure at the nascent site of cytokinesis determines the timing and position of division in most bacteria. This FtsZ ring serves as a scaffold for recruitment of other proteins into a mature division-competent structure permitting membrane constriction and septal cell wall synthesis. Here, we show that expression of the predicted 9.3-kDa gp56 of SP01 inhibits later stages of B. subtilis cell division without altering FtsZ ring assembly. Green fluorescent protein-tagged gp56 localizes to the membrane at the site of division. While its localization does not interfere with recruitment of early division proteins, gp56 interferes with the recruitment of late division proteins, including Pbp2b and FtsW. Imaging of cells with specific division components deleted or depleted and two-hybrid analyses suggest that gp56 localization and activity depend on its interaction with FtsL. Together, these data support a model in which gp56 interacts with a central part of the division machinery to disrupt late recruitment of the division proteins involved in septal cell wall synthesis.IMPORTANCE Studies over the past decades have identified bacteriophage-encoded factors that interfere with host cell shape or cytokinesis during viral infection. The phage factors causing cell filamentation that have been investigated to date all act by targeting FtsZ, the conserved prokaryotic tubulin homolog that composes the cytokinetic ring in most bacteria and some groups of archaea. However, the mechanisms of several phage factors that inhibit cytokinesis, including gp56 of bacteriophage SP01 of Bacillus subtilis, remain unexplored. Here, we show that, unlike other published examples of phage inhibition of cytokinesis, gp56 blocks B. subtilis cell division without targeting FtsZ. Rather, it utilizes the assembled FtsZ cytokinetic ring to localize to the division machinery and to block recruitment of proteins needed for septal cell wall synthesis.

scholarly journals The heat-shock response of Listeria monocytogenes comprises genes involved in heat shock, cell division, cell wall synthesis, and the SOS response

Microbiology ◽  
2007 ◽  
Vol 153 (10) ◽  
pp. 3593-3607 ◽  
Author(s):  
Stijn van der Veen ◽  
Torsten Hain ◽  
Jeroen A. Wouters ◽  
Hamid Hossain ◽  
Willem M. de Vos ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Division ◽  
Listeria Monocytogenes ◽  
Heat Shock ◽  
Heat Shock Response ◽  
Sos Response ◽  
Cell Wall Synthesis ◽  
Shock Cell ◽  
Shock Response ◽  
Division Cell

scholarly journals Cell Shape and Antibiotic Resistance Are Maintained by the Activity of Multiple FtsW and RodA Enzymes in Listeria monocytogenes

mBio ◽  
2019 ◽  
Vol 10 (4) ◽  
Author(s):  
Jeanine Rismondo ◽  
Sven Halbedel ◽  
Angelika Gründling
Keyword(s):  
Cell Wall ◽  
Antibiotic Resistance ◽  
Immune System ◽  
Cell Division ◽  
Listeria Monocytogenes ◽  
Antimicrobial Agents ◽  
Protein Complexes ◽  
Inducible Promoter ◽  
Human Pathogen ◽  
Content Type

ABSTRACT Rod-shaped bacteria have two modes of peptidoglycan synthesis: lateral synthesis and synthesis at the cell division site. These two processes are controlled by two macromolecular protein complexes, the elongasome and divisome. Recently, it has been shown that the Bacillus subtilis RodA protein, which forms part of the elongasome, has peptidoglycan glycosyltransferase activity. The cell division-specific RodA homolog FtsW fulfils a similar role at the divisome. The human pathogen Listeria monocytogenes carries genes that encode up to six FtsW/RodA homologs; however, their functions have not yet been investigated. Analysis of deletion and depletion strains led to the identification of the essential cell division-specific FtsW protein, FtsW1. Interestingly, L. monocytogenes carries a gene that encodes a second FtsW protein, FtsW2, which can compensate for the lack of FtsW1, when expressed from an inducible promoter. L. monocytogenes also possesses three RodA homologs, RodA1, RodA2, and RodA3, and their combined absence is lethal. Cells of a rodA1 rodA3 double mutant are shorter and have increased antibiotic and lysozyme sensitivity, probably due to a weakened cell wall. Results from promoter activity assays revealed that expression of rodA3 and ftsW2 is induced in the presence of antibiotics targeting penicillin binding proteins. Consistent with this, a rodA3 mutant was more susceptible to the β-lactam antibiotic cefuroxime. Interestingly, overexpression of RodA3 also led to increased cefuroxime sensitivity. Our study highlights that L. monocytogenes genes encode a multitude of functional FtsW and RodA enzymes to produce its rigid cell wall and that their expression needs to be tightly regulated to maintain growth, cell division, and antibiotic resistance. IMPORTANCE The human pathogen Listeria monocytogenes is usually treated with high doses of β-lactam antibiotics, often combined with gentamicin. However, these antibiotics only act bacteriostatically on L. monocytogenes, and the immune system is needed to clear the infection. Therefore, individuals with a compromised immune system are at risk to develop a severe form of Listeria infection, which can be fatal in up to 30% of cases. The development of new strategies to treat Listeria infections is necessary. Here we show that the expression of some of the FtsW and RodA enzymes of L. monocytogenes is induced by the presence of β-lactam antibiotics, and the combined absence of these enzymes makes bacteria more susceptible to this class of antibiotics. The development of antimicrobial agents that inhibit the activity or production of FtsW and RodA enzymes might therefore help to improve the treatment of Listeria infections and thereby lead to a reduction in mortality.

scholarly journals Suppressor Mutations Linking gpsB with the First Committed Step of Peptidoglycan Biosynthesis in Listeria monocytogenes

2016 ◽  
Vol 199 (1) ◽  
Author(s):  
Jeanine Rismondo ◽  
Jennifer K. Bender ◽  
Sven Halbedel
Keyword(s):  
Cell Wall ◽  
Cell Division ◽  
Listeria Monocytogenes ◽  
Cell Wall Biosynthesis ◽  
Human Pathogen ◽  
Penicillin Binding Protein ◽  
Content Type ◽  
Peptidoglycan Biosynthesis ◽  
Cell Division Protein ◽  
Precursor Molecules

ABSTRACT The cell division protein GpsB is a regulator of the penicillin binding protein A1 (PBP A1) in the Gram-positive human pathogen Listeria monocytogenes. Penicillin binding proteins mediate the last two steps of peptidoglycan biosynthesis as they polymerize and cross-link peptidoglycan strands, the main components of the bacterial cell wall. It is not known what other processes are controlled by GpsB. L. monocytogenes gpsB mutants are unable to grow at 42°C, but we observed that spontaneous suppressors correcting this defect arise on agar plates with high frequency. We here describe a first set of gpsB suppressors that mapped to the clpC and murZ genes. While ClpC is the ATPase component of the Clp protease, MurZ is a paralogue of the listerial UDP–N-acetylglucosamine (UDP-GlcNAc) 1-carboxyvinyltransferase MurA. Both enzymes catalyze the enolpyruvyl transfer from phosphoenolpyruvate to UDP-GlcNAc, representing the first committed step of peptidoglycan biosynthesis. We confirmed that clean deletion of the clpC or murZ gene suppressed the ΔgpsB phenotype. It turned out that the absence of either gene leads to accumulation of MurA, and we show that artificial overexpression of MurA alone was sufficient for suppression. Inactivation of other UDP-GlcNAc-consuming pathways also suppressed the heat-sensitive growth of the ΔgpsB mutant, suggesting that an increased influx of precursor molecules into peptidoglycan biosynthesis can compensate for the lack of GpsB. Our results support a model according to which PBP A1 becomes misregulated and thus toxic in the absence of GpsB due to unproductive consumption of cell wall precursor molecules. IMPORTANCE The late cell division protein GpsB is important for cell wall biosynthesis in Gram-positive bacteria. GpsB of the human pathogen L. monocytogenes interacts with one of the key enzymes of this pathway, penicillin binding protein A1 (PBP A1), and influences its activity. PBP A1 catalyzes the last two steps of cell wall biosynthesis, but it is unknown how GpsB controls PBP A1. We observed that a L. monocytogenes gpsB mutant forms spontaneous suppressors and have mapped their mutations to genes mediating and influencing the first step of cell wall biosynthesis, likely stimulating the influx of metabolites into this pathway. We assume that GpsB is important to ensure productive incorporation of cell wall precursors into the peptidoglycan sacculus by PBP A1.

scholarly journals Interplay between Antibiotic Efficacy and Drug-Induced Lysis Underlies Enhanced Biofilm Formation at Subinhibitory Drug Concentrations

2017 ◽  
Vol 62 (1) ◽  
Author(s):  
Wen Yu ◽  
Kelsey M. Hallinen ◽  
Kevin B. Wood
Keyword(s):  
Cell Wall ◽  
Biofilm Formation ◽  
Cell Lysis ◽  
Living Cells ◽  
Cell Wall Synthesis ◽  
Drug Induced ◽  
Trade Off ◽  
Content Type ◽  
Subinhibitory Concentrations ◽  
Antibiotic Efficacy

ABSTRACTSubinhibitory concentrations of antibiotics have been shown to enhance biofilm formation in multiple bacterial species. While antibiotic exposure has been associated with modulated expression of many biofilm-related genes, the mechanisms of drug-induced biofilm formation remain a focus of ongoing research efforts and may vary significantly across species. In this work, we investigate antibiotic-induced biofilm formation inEnterococcus faecalis, a leading cause of nosocomial infections. We show that biofilm formation is enhanced by subinhibitory concentrations of cell wall synthesis inhibitors but not by inhibitors of protein, DNA, folic acid, or RNA synthesis. Furthermore, enhanced biofilm is associated with increased cell lysis, increases in extracellular DNA (eDNA) levels, and increases in the density of living cells in the biofilm. In addition, we observe similar enhancement of biofilm formation when cells are treated with nonantibiotic surfactants that induce cell lysis. These findings suggest that antibiotic-induced biofilm formation is governed by a trade-off between drug toxicity and the beneficial effects of cell lysis. To understand this trade-off, we developed a simple mathematical model that predicts changes in antibiotic-induced biofilm formation due to external perturbations, and we verified these predictions experimentally. Specifically, we demonstrate that perturbations that reduce eDNA (DNase treatment) or decrease the number of living cells in the planktonic phase (a second antibiotic) decrease biofilm induction, while chemical inhibitors of cell lysis increase relative biofilm induction and shift the peak to higher antibiotic concentrations. Overall, our results offer experimental evidence linking cell wall synthesis inhibitors, cell lysis, increased eDNA levels, and biofilm formation inE. faecaliswhile also providing a predictive quantitative model that sheds light on the interplay between cell lysis and antibiotic efficacy in developing biofilms.

scholarly journals Draft Whole-Genome Sequences of Seven Listeria monocytogenes Strains with Variations in Virulence and Stress Responses

2018 ◽  
Vol 7 (13) ◽  
Author(s):  
Yanhong Liu ◽  
Aixia Xu ◽  
Pina M. Fratamico ◽  
Christopher H. Sommers ◽  
Luca Rotundo ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Responses ◽  
Draft Genome ◽  
Foodborne Pathogen ◽  
Whole Genome ◽  
Genome Sequences ◽  
Content Type

Listeria monocytogenes is an important foodborne pathogen that causes listeriosis. Here, we report the draft genome sequences of seven L. monocytogenes strains isolated from food, environmental, and clinical sources.

scholarly journals Cell shape and antibiotic resistance is maintained by the activity of multiple FtsW and RodA enzymes inListeria monocytogenes

2019 ◽  
Author(s):  
Jeanine Rismondo ◽  
Sven Halbedel ◽  
Angelika Gründling
Keyword(s):  
Cell Wall ◽  
Antibiotic Resistance ◽  
Immune System ◽  
Cell Division ◽  
Listeria Monocytogenes ◽  
Protein Complexes ◽  
Inducible Promoter ◽  
Severe Form ◽  
Human Pathogen ◽  
Division Site

AbstractRod-shaped bacteria have two modes of peptidoglycan synthesis: lateral synthesis and synthesis at the cell division site. These two processes are controlled by two macromolecular protein complexes, the elongasome and divisome. Recently, it has been shown that theBacillus subtilisRodA protein, which forms part of the elongasome, has peptidoglycan glycosyltransferase activity. The cell division specific RodA homolog FtsW fulfils a similar role at the divisome. The human pathogenListeria monocytogenesencodes up to six FtsW/RodA homologs, however their functions have not yet been investigated. Analysis of deletion and depletion strains led to the identification of the essential cell division-specific FtsW protein, FtsW1. Interestingly,L. monocytogenesencodes a second FtsW protein, FtsW2, which can compensate for the lack of FtsW1, when expressed from an inducible promoter.L. monocytogenesalso possesses three RodA homologs, RodA1, RodA2 and RodA3 and their combined absence is lethal. Cells of arodA1/rodA3double mutant are shorter and have increased antibiotic and lysozyme sensitivity, probably due to a weakened cell wall. Results from promoter activity assays revealed that expression ofrodA3andftsW2is induced in the presence of antibiotics targeting penicillin binding proteins. Consistent with this, arodA3mutant was more susceptible to the β-lactam antibiotic cefuroxime. Interestingly, overexpression of RodA3 also led to increased cefuroxime sensitivity. Our study highlights thatL. monocytogenesencodes a multitude of functional FtsW and RodA enzymes to produce its rigid cell wall and that their expression needs to be tightly regulated to maintain growth, cell division and antibiotic resistance.ImportanceThe human pathogenListeria monocytogenesis usually treated with high doses of β-lactam antibiotics, often combined with gentamicin. However, these antibiotics only act bacteriostatically onL. monocytogenesand the immune system is needed to clear the infection. Therefore, individuals with a compromised immune system are at risk to develop a severe form ofListeriainfection, which can be fatal in up to 30% of cases. The development of new strategies to treatListeriainfections is therefore necessary. Here we show that the expression of some of the FtsW and RodA enzymes ofL. monocytogenesis induced by the presence of β-lactam antibiotics and their combined absence makes bacteria more susceptible to this class of antibiotics. The development of antimicrobials that inhibit the activity or production of FtsW/RodA enzymes might therefore help to improve the treatment ofListeriainfections and thereby lead to a reduction in mortality.

scholarly journals EslB is required for cell wall biosynthesis and modification in Listeria monocytogenes

2020 ◽  
Author(s):  
Jeanine Rismondo ◽  
Lisa M. Schulz ◽  
Maria Yacoub ◽  
Ashima Wadhawan ◽  
Michael Hoppert ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Division ◽  
Listeria Monocytogenes ◽  
Abc Transporter ◽  
Cell Lysis ◽  
Growth Conditions ◽  
Cell Wall Integrity ◽  
Growth Defect ◽  
Cell Wall Biosynthesis ◽  
High Concentrations

Lysozyme is an important component of the innate immune system. It functions by hydrolysing the peptidoglycan (PG) layer of bacteria. The human pathogen Listeria monocytogenes is intrinsically lysozyme resistant. The peptidoglycan N-deacetylase PgdA and O-acetyltransferase OatA are two known factors contributing to its lysozyme resistance. Furthermore, it was shown that the absence of components of an ABC transporter, here referred to as EslABC, leads to reduced lysozyme resistance. How its activity is linked to lysozyme resistance is still unknown. To investigate this further, a strain with a deletion in eslB, coding for a membrane component of the ABC transporter, was constructed in L. monocytogenes strain 10403S. The eslB mutant showed a 40-fold reduction in the minimal inhibitory concentration to lysozyme. Analysis of the PG structure revealed that the eslB mutant produced PG with reduced levels of O-acetylation. Using growth and autolysis assays, we show that the absence of EslB manifests in a growth defect in media containing high concentrations of sugars and increased endogenous cell lysis. A thinner PG layer produced by the eslB mutant under these growth conditions might explain these phenotypes. Furthermore, the eslB mutant had a noticeable cell division defect and formed elongated cells. Microscopy analysis revealed that an early cell division protein still localized in the eslB mutant indicating that a downstream process is perturbed. Based on our results, we hypothesize that EslB affects the biosynthesis and modification of the cell wall in L. monocytogenes and is thus important for the maintenance of cell wall integrity. IMPORTANCE The ABC transporter EslABC is associated with the intrinsic lysozyme resistance of Listeria monocytogenes. However, the exact role of the transporter in this process and in the physiology of L. monocytogenes is unknown. Using different assays to characterize an eslB deletion strain, we found that the absence of EslB not only affects lysozyme resistance, but also endogenous cell lysis, cell wall biosynthesis, cell division and the ability of the bacterium to grow in media containing high concentrations of sugars. Our results indicate that EslB is by a yet unknown mechanism an important determinant for cell wall integrity in L. monocytogenes.

scholarly journals Interaction of Platelets and Anidulafungin against Aspergillus fumigatus

2012 ◽  
Vol 57 (1) ◽  
pp. 626-628 ◽  
Author(s):  
Susanne Perkhofer ◽  
Barbara Striessnig ◽  
Bettina Sartori ◽  
Barbara Hausott ◽  
Helmut W. Ott ◽  
...  
Keyword(s):  
Cell Wall ◽  
Aspergillus Fumigatus ◽  
Untreated Control ◽  
Germination Rate ◽  
Human Platelets ◽  
Cell Wall Synthesis ◽  
Content Type

ABSTRACTThe combination of platelets and anidulafungin at 0.03 μg/ml significantly (P< 0.05) reduced the germination rate and hyphal elongation inAspergillus fumigatuscompared to those with either anidulafungin only or an untreated control. Platelets decreased the expression of thefksgene, which plays an important role in cell wall synthesis. Our results suggest that human platelets plus anidulafungin might contribute to defense againstA. fumigatus.

scholarly journals Genetic Dissection of DivIVA Functions in Listeria monocytogenes

2017 ◽  
Vol 199 (24) ◽  
Author(s):  
Karan Gautam Kaval ◽  
Samuel Hauf ◽  
Jeanine Rismondo ◽  
Birgitt Hahn ◽  
Sven Halbedel
Keyword(s):  
Cell Division ◽  
Listeria Monocytogenes ◽  
Protein Interactions ◽  
Site Selection ◽  
Genetic Dissection ◽  
Flagellar Motility ◽  
Content Type ◽  
Division Site ◽  
Interaction Partners ◽  
Species Specific

ABSTRACT DivIVA is a membrane binding protein that clusters at curved membrane regions, such as the cell poles and the membrane invaginations occurring during cell division. DivIVA proteins recruit many other proteins to these subcellular sites through direct protein-protein interactions. DivIVA-dependent functions are typically associated with cell growth and division, even though species-specific differences in the spectrum of DivIVA functions and their causative interaction partners exist. DivIVA from the Gram-positive human pathogen Listeria monocytogenes has at least three different functions. In this bacterium, DivIVA is required for precise positioning of the septum at midcell, it contributes to the secretion of autolysins required for the breakdown of peptidoglycan at the septum after the completion of cell division, and it is essential for flagellar motility. While the DivIVA interaction partners for control of division site selection are well established, the proteins connecting DivIVA with autolysin secretion or swarming motility are completely unknown. We set out to identify divIVA alleles in which these three DivIVA functions could be separated, since the question of the degree to which the three functions of L. monocytogenes DivIVA are interlinked could not be answered before. Here, we identify such alleles, and our results show that division site selection, autolysin secretion, and swarming represent three discrete pathways that are independently influenced by DivIVA. These findings provide the required basis for the identification of DivIVA interaction partners controlling autolysin secretion and swarming in the future. IMPORTANCE DivIVA of the pathogenic bacterium Listeria monocytogenes is a central scaffold protein that influences at least three different cellular processes, namely, cell division, protein secretion, and bacterial motility. How DivIVA coordinates these rather unrelated processes is not known. We here identify variants of L. monocytogenes DivIVA, in which these functions are separated from each other. These results have important implications for the models explaining how DivIVA interacts with other proteins.

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