scholarly journals Truncation in the core oligosaccharide of lipopolysaccharide affects flagella-mediated motility in Pseudomonas aeruginosa PAO1 via modulation of cell surface attachment

Microbiology ◽  
2009 ◽  
Vol 155 (10) ◽  
pp. 3449-3460 ◽  
Author(s):  
Theresa Lindhout ◽  
Peter C. Y. Lau ◽  
Dyanne Brewer ◽  
Joseph S. Lam
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Surface ◽  
Bacterial Species ◽  
Soft Agar ◽  
Wild Type ◽  
Core Oligosaccharide ◽  
Swarming Motility ◽  
Surface Attachment ◽  
Abiotic Surfaces ◽  
Isogenic Mutants

In many Gram-negative bacterial species, rough strains producing truncated lipopolysaccharide (LPS) generally exhibit defects in motility compared with smooth strains. However, the role that LPS plays in bacterial motility is not well understood. The goal of this study was to examine the relationship between LPS defects and motility of Pseudomonas aeruginosa. P. aeruginosa wild-type strain PAO1 and three isogenic mutants with defects in the rmlC, migA and wapR genes and producing truncated core oligosaccharide were investigated in terms of motility, attachment to glass and flagella expression. Compared with the wild-type, the three mutants showed significant retardation in both swarming motility on 0.5 % soft-agar plates and swimming motility on 0.3 % soft-agar plates. Moreover, attachment to abiotic surfaces was observed to be stronger in these mutants. The assembly of flagella appeared to be intact in these strains and the ability of individual cells to swim was unaffected. Flagellin proteins prepared from mutants rmlC and rmd, defective in the production of TDP-l-rhamnose and GDP-d-rhamnose, respectively, were compared and a change in molecular mass was observed only in the rmlC mutant. These data indicated that l-rhamnose, and not its enantiomer, d-rhamnose, is incorporated into the flagellin glycan of P. aeruginosa PAO1. The nucleotide-activated sugar precursor TDP-l-rhamnose is therefore shared between LPS biosynthesis and flagellin glycosylation in P. aeruginosa PAO1. Our results suggest that although biochemical precursors are shared by LPS and flagellin glycan biosynthesis, LPS truncations probably alter flagella-mediated motility in P. aeruginosa by modulating cell-surface attachment but not flagella synthesis.

scholarly journals FlhF Is Required for Swimming and Swarming in Pseudomonas aeruginosa

2006 ◽  
Vol 188 (19) ◽  
pp. 6995-7004 ◽  
Author(s):  
Thomas S. Murray ◽  
Barbara I. Kazmierczak
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Surface ◽  
Signal Recognition Particle ◽  
Signal Recognition ◽  
Wild Type ◽  
Liquid Media ◽  
Swarming Motility ◽  
Flagellar Assembly ◽  
Swarming Behavior ◽  
Flagellar Genes

ABSTRACT FlhF is a signal recognition particle-like protein present in monotrichous bacteria. The loss of FlhF in various bacteria results in decreased transcription of class II, III, or IV flagellar genes, leads to diminished or absent motility, and results in the assembly of flagella at nonpolar locations on the cell surface. In this work, we demonstrate that the loss of FlhF results in defective swimming and swarming motility of Pseudomonas aeruginosa. The FlhF protein localizes to the flagellar pole; in the absence of FlhF, flagellar assembly occurs but is no longer restricted to the pole. ΔflhF bacteria swim at lower velocities than wild-type bacteria in liquid media and can no longer swarm when assayed under standard swarming conditions (0.5% agar). However, ΔflhF bacteria regain swarming behavior when plated on 0.3% agar. ΔflhF organisms show decreased transcription and expression of flagellin (FliC) both in liquid media and on swarming plates compared to wild-type bacteria. However, changes in flagellin expression do not explain the different motility patterns observed for ΔflhF bacteria. Instead, the aberrant placement of flagella in ΔflhF bacteria may reduce their ability to move this rod-shaped organism effectively.

scholarly journals Motility and Chemotaxis in Agrobacterium tumefaciens Surface Attachment and Biofilm Formation

2007 ◽  
Vol 189 (22) ◽  
pp. 8005-8014 ◽  
Author(s):  
Peter M. Merritt ◽  
Thomas Danhorn ◽  
Clay Fuqua
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Wild Type ◽  
Swarming Motility ◽  
Surface Attachment ◽  
Directed Motility ◽  
Suppressor Mutants ◽  
Static Conditions ◽  
Specific Deletion

ABSTRACT Bacterial motility mechanisms, including swimming, swarming, and twitching, are known to have important roles in biofilm formation, including colonization and the subsequent expansion into mature structured surface communities. Directed motility requires chemotaxis functions that are conserved among many bacterial species. The biofilm-forming plant pathogen Agrobacterium tumefaciens drives swimming motility by utilizing a small group of flagella localized to a single pole or the subpolar region of the cell. There is no evidence for twitching or swarming motility in A. tumefaciens. Site-specific deletion mutations that resulted in either aflagellate, flagellated but nonmotile, or flagellated but nonchemotactic A. tumefaciens derivatives were examined for biofilm formation under static and flowing conditions. Nonmotile mutants were significantly deficient in biofilm formation under static conditions. Under flowing conditions, however, the aflagellate mutant rapidly formed aberrantly dense, tall biofilms. In contrast, a nonmotile mutant with unpowered flagella was clearly debilitated for biofilm formation relative to the wild type. A nontumbling chemotaxis mutant was only weakly affected with regard to biofilm formation under nonflowing conditions but was notably compromised in flow, generating less adherent biomass than the wild type, with a more dispersed cellular arrangement. Extragenic suppressor mutants of the chemotaxis-impaired, straight-swimming phenotype were readily isolated from motility agar plates. These mutants regained tumbling at a frequency similar to that of the wild type. Despite this phenotype, biofilm formation by the suppressor mutants in static cultures was significantly deficient. Under flowing conditions, a representative suppressor mutant manifested a phenotype similar to yet distinct from that of its nonchemotactic parent.

scholarly journals Effect of Nitroxides on Swarming Motility and Biofilm Formation, Multicellular Behaviors in Pseudomonas aeruginosa

2013 ◽  
Vol 57 (10) ◽  
pp. 4877-4881 ◽  
Author(s):  
César de la Fuente-Núñez ◽  
Fany Reffuveille ◽  
Kathryn E. Fairfull-Smith ◽  
Robert E. W. Hancock
Keyword(s):  
Nitric Oxide ◽  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Wild Type ◽  
Planktonic Cells ◽  
Swarming Motility ◽  
Content Type ◽  
Exogenous Addition ◽  
Biofilm Dispersion ◽  
Dispersal Activity

ABSTRACTThe ability of nitric oxide (NO) to induce biofilm dispersion has been well established. Here, we investigated the effect of nitroxides (sterically hindered nitric oxide analogues) on biofilm formation and swarming motility inPseudomonas aeruginosa. A transposon mutant unable to produce nitric oxide endogenously (nirS) was deficient in swarming motility relative to the wild type and the complemented strain. Moreover, expression of thenirSgene was upregulated by 9.65-fold in wild-type swarming cells compared to planktonic cells. Wild-type swarming levels were substantially restored upon the exogenous addition of nitroxide containing compounds, a finding consistent with the hypothesis that NO is necessary for swarming motility. Here, we showed that nitroxides not only mimicked the dispersal activity of NO but also prevented biofilms from forming in flow cell chambers. In addition, anirStransposon mutant was deficient in biofilm formation relative to the wild type and the complemented strain, thus implicating NO in the formation of biofilms. Intriguingly, despite its stand-alone action in inhibiting biofilm formation and promoting dispersal, a nitroxide partially restored the ability of anirSmutant to form biofilms.

scholarly journals Rhamnolipids Modulate Swarming Motility Patterns of Pseudomonas aeruginosa

2005 ◽  
Vol 187 (21) ◽  
pp. 7351-7361 ◽  
Author(s):  
Nicky C. Caiazza ◽  
Robert M. Q. Shanks ◽  
G. A. O'Toole
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Genetic Analysis ◽  
Group Behavior ◽  
Biosurfactant Production ◽  
Wild Type ◽  
Swarming Motility

ABSTRACT Pseudomonas aeruginosa is capable of twitching, swimming, and swarming motility. The latter form of translocation occurs on semisolid surfaces, requires functional flagella and biosurfactant production, and results in complex motility patterns. From the point of inoculation, bacteria migrate as defined groups, referred to as tendrils, moving in a coordinated manner capable of sensing and responding to other groups of cells. We were able to show that P. aeruginosa produces extracellular factors capable of modulating tendril movement, and genetic analysis revealed that modulation of these movements was dependent on rhamnolipid biosynthesis. An rhlB mutant (deficient in mono- and dirhamnolipid production) and an rhlC mutant (deficient in dirhamnolipid production) exhibited altered swarming patterns characterized by irregularly shaped tendrils. In addition, agar supplemented with rhamnolipid-containing spent supernatant inhibited wild-type (WT) swarming, whereas agar supplemented with spent supernatant from mutants that do not make rhamnolipids had no effect on WT P. aeruginosa swarming. Addition of purified rhamnolipids to swarming medium also inhibited swarming motility of the WT strain. We also show that a sadB mutant does not sense and/or respond to other groups of swarming cells and this mutant was capable of swarming on media supplemented with rhamnolipid-containing spent supernatant or purified rhamnolipids. The abilities to produce and respond to rhamnolipids in the context of group behavior are discussed.

scholarly journals Heterogeneity in surface sensing produces a division of labor in Pseudomonas aeruginosa populations

2019 ◽  
Author(s):  
Catherine R. Armbruster ◽  
Calvin K. Lee ◽  
Jessica Parker-Gilham ◽  
Jaime de Anda ◽  
Aiguo Xia ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Division Of Labor ◽  
Bacterial Species ◽  
Biofilm Growth ◽  
Surface Attachment ◽  
Sensing Systems ◽  
Current Thinking ◽  
Surface Exploration ◽  
Surface Sensing ◽  
Second Messenger Signaling

AbstractThe second messenger signaling molecule cyclic diguanylate monophosphate (c-di-GMP) drives the transition from planktonic to biofilm growth in many bacterial species. Pseudomonas aeruginosa has two surface sensing systems that produce c-di-GMP in response to surface adherence. The current thinking in the field is that once cells attach to a surface, they uniformly respond with elevated c-di-GMP. Here, we describe how the Wsp system generates heterogeneity in surface sensing, resulting in two physiologically distinct subpopulations of cells. One subpopulation has elevated c-di-GMP and produces biofilm matrix, serving as the founders of initial microcolonies. The other subpopulation has low c-di-GMP and engages in surface motility, allowing for exploration of the surface. We also show that this heterogeneity strongly correlates to surface behavior for descendent cells. Together, our results suggest that after surface attachment, P. aeruginosa engages in a division of labor that persists across generations, accelerating early biofilm formation and surface exploration.

scholarly journals Retarded swarming motility in Bacillus subtilis NRS-762 and Pseudomonas aeruginosa PRD-10

2018 ◽  
Author(s):  
Wenfa Ng
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Bacillus Subtilis ◽  
Bacterial Species ◽  
Nutritional Stress ◽  
Cellular Level ◽  
Bacterial Cells ◽  
Loss Of Function ◽  
Swarming Motility ◽  
Bacteria Growth ◽  
Solid Media

Coping with nutritional stress is essential for cell survival, of which many strategies at the cellular level lend support for ensuring the survival of the population at a particular habitat. One postulated mechanism is swarming motility in bacterial cells, where, upon depletion of nutrients at a locale, cells would coordinate their movement, synthesize more flagella, and secrete lubricants for moving rapidly across surfaces in search for food. Known to engage in swarming motility, Bacillus subtilis and Pseudomonas aeruginosa are two common bacterial species with versatile metabolism that use the motility mode to colonize new habitats with more favourable environmental and nutritional conditions. However, experimental observations of bacteria growth on a variety of agar media revealed that B. subtilis NRS-762 (ATCC 8473) and P. aeruginosa PRD-10 (ATCC 15442) exhibited retarded swarming motility upon entry into stationary phase on solid media. Specifically, B. subtilis NRS-762 colonies exhibited round, wrinkled morphologies compared to complex filamented swarming patterns common in strains able to engage in swarming motility. On the other hand, P. aeruginosa PRD-10 colonies were round, mucoid, and expanded outwards from the colony centre without extending filaments from the centre; thereby, indicating retarded swarming motility. Thus, impaired cellular machinery for swarming motility or mutated and deleted genes likely account for observed retarded swarming motility in B. subtilis NRS-762 and P. aeruginosa PRD-10. More importantly, observations of small filaments extending radially from an expanded colony of P. aeruginosa PRD-10 grown on minimal salts medium supplemented with yeast extract highlighted possible loss of function of effector molecules that transmit cellular decision at swarming motility into movement, while sensory mechanisms feeding into the motility mechanism remained intact. More broadly, observations of impaired swarming motility in B. subtilis NRS-762 and P. aeruginosa PRD-10 in two species otherwise endowed with the motility mode highlighted that additional triggers for swarming motility are likely present, and the motility mode may have been evolutionary selected for other functions in addition to foraging for food in times of nutritional stress.

scholarly journals Heterogeneity in surface sensing suggests a division of labor in Pseudomonas aeruginosa populations

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Catherine R Armbruster ◽  
Calvin K Lee ◽  
Jessica Parker-Gilham ◽  
Jaime de Anda ◽  
Aiguo Xia ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Division Of Labor ◽  
Bacterial Species ◽  
Biofilm Growth ◽  
Surface Attachment ◽  
Sensing Systems ◽  
Current Thinking ◽  
Surface Exploration ◽  
Surface Sensing ◽  
Second Messenger Signaling

The second messenger signaling molecule cyclic diguanylate monophosphate (c-di-GMP) drives the transition between planktonic and biofilm growth in many bacterial species. Pseudomonas aeruginosa has two surface sensing systems that produce c-di-GMP in response to surface adherence. Current thinking in the field is that once cells attach to a surface, they uniformly respond by producing c-di-GMP. Here, we describe how the Wsp system generates heterogeneity in surface sensing, resulting in two physiologically distinct subpopulations of cells. One subpopulation has elevated c-di-GMP and produces biofilm matrix, serving as the founders of initial microcolonies. The other subpopulation has low c-di-GMP and engages in surface motility, allowing for exploration of the surface. We also show that this heterogeneity strongly correlates to surface behavior for descendent cells. Together, our results suggest that after surface attachment, P. aeruginosa engages in a division of labor that persists across generations, accelerating early biofilm formation and surface exploration.

scholarly journals Global Analysis of Cellular Factors and Responses Involved in Pseudomonas aeruginosa Resistance to Arsenite

2005 ◽  
Vol 187 (14) ◽  
pp. 4853-4864 ◽  
Author(s):  
Kislay Parvatiyar ◽  
Eyad M. Alsabbagh ◽  
Urs A. Ochsner ◽  
Michelle A. Stegemeyer ◽  
Alan G. Smulian ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pseudomonas Aeruginosa ◽  
Glutathione Reductase ◽  
Type Strain ◽  
Wild Type Strain ◽  
Storage Proteins ◽  
Global Analysis ◽  
Wild Type ◽  
Isogenic Mutants ◽  
The Impact

ABSTRACT The impact of arsenite [As(III)] on several levels of cellular metabolism and gene regulation was examined in Pseudomonas aeruginosa. P. aeruginosa isogenic mutants devoid of antioxidant enzymes or defective in various metabolic pathways, DNA repair systems, metal storage proteins, global regulators, or quorum sensing circuitry were examined for their sensitivity to As(III). Mutants lacking the As(III) translocator (ArsB), superoxide dismutase (SOD), catabolite repression control protein (Crc), or glutathione reductase (Gor) were more sensitive to As(III) than wild-type bacteria. The MICs of As(III) under aerobic conditions were 0.2, 0.3, 0.8, and 1.9 mM for arsB, sodA sodB, crc, and gor mutants, respectively, and were 1.5- to 13-fold less than the MIC for the wild-type strain. A two-dimensional gel/matrix-assisted laser desorption ionization-time of flight analysis of As(III)-treated wild-type bacteria showed significantly (>40-fold) increased levels of a heat shock protein (IbpA) and a putative allo-threonine aldolase (GlyI). Smaller increases (up to 3.1-fold) in expression were observed for acetyl-coenzyme A acetyltransferase (AtoB), a probable aldehyde dehydrogenase (KauB), ribosomal protein L25 (RplY), and the probable DNA-binding stress protein (PA0962). In contrast, decreased levels of a heme oxygenase (HemO/PigA) were found upon As(III) treatment. Isogenic mutants were successfully constructed for six of the eight genes encoding the aforementioned proteins. When treated with sublethal concentrations of As(III), each mutant revealed a marginal to significant lag period prior to resumption of apparent normal growth compared to that observed in the wild-type strain. Our results suggest that As(III) exposure results in an oxidative stress-like response in P. aeruginosa, although activities of classic oxidative stress enzymes are not increased. Instead, relief from As(III)-based oxidative stress is accomplished from the collective activities of ArsB, glutathione reductase, and the global regulator Crc. SOD appears to be involved, but its function may be in the protection of superoxide-sensitive sulfhydryl groups.

scholarly journals Cluster II che Genes from Pseudomonas aeruginosa Are Required for an Optimal Chemotactic Response

2002 ◽  
Vol 184 (16) ◽  
pp. 4374-4383 ◽  
Author(s):  
Abel Ferrández ◽  
Andrew C. Hawkins ◽  
Douglas T. Summerfield ◽  
Caroline S. Harwood
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Gene Clusters ◽  
Soft Agar ◽  
Chemotactic Response ◽  
Wild Type ◽  
E Coli ◽  
Chemotaxis Proteins ◽  
Complete Set ◽  
K 12

ABSTRACT Pseudomonas aeruginosa, a γ-proteobacterium, is motile by means of a single polar flagellum and is chemotactic to a variety of organic compounds and phosphate. P. aeruginosa has multiple homologues of Escherichia coli chemotaxis genes that are organized into five gene clusters. Previously, it was demonstrated that genes in cluster I and cluster V are essential for chemotaxis. A third cluster (cluster II) contains a complete set of che genes, as well as two genes, mcpA and mcpB, encoding methyl-accepting chemotaxis proteins. Mutations were constructed in several of the cluster II che genes and in the mcp genes to examine their possible contributions to P. aeruginosa chemotaxis. A cheB2 mutant was partially impaired in chemotaxis in soft-agar swarm plate assays. Providing cheB2 in trans complemented this defect. Further, overexpression of CheB2 restored chemotaxis to a completely nonchemotactic, cluster I, cheB-deficient strain to near wild-type levels. An mcpA mutant was defective in chemotaxis in media that were low in magnesium. The defect could be relieved by the addition of magnesium to the swarm plate medium. An mcpB mutant was defective in chemotaxis when assayed in dilute rich soft-agar swarm medium or in minimal-medium swarm plates containing any 1 of 60 chemoattractants. The mutant phenotype could be complemented by the addition of mcpB in trans. Overexpression of either McpA or McpB in P. aeruginosa or Escherichia coli resulted in impairment of chemotaxis, and these cells had smooth-swimming phenotypes when observed under the microscope. Expression of P. aeruginosa cheA2, cheB2, or cheW2 in E. coli K-12 completely disrupted wild-type chemotaxis, while expression of cheY2 had no effect. These results indicate that che cluster II genes are expressed in P. aeruginosa and are required for an optimal chemotactic response.

scholarly journals Searching for the Secret of Stickiness: How Biofilms Adhere to Surfaces

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhaowei Jiang ◽  
Thomas Nero ◽  
Sampriti Mukherjee ◽  
Rich Olson ◽  
Jing Yan
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Cell Surface ◽  
Dynamic Process ◽  
Population Expansion ◽  
Bacterial Biofilms ◽  
Surface Attachment ◽  
Matrix Production ◽  
Main Factors ◽  
Novel Approaches

Bacterial biofilms are communities of cells enclosed in an extracellular polymeric matrix in which cells adhere to each other and to foreign surfaces. The development of a biofilm is a dynamic process that involves multiple steps, including cell-surface attachment, matrix production, and population expansion. Increasing evidence indicates that biofilm adhesion is one of the main factors contributing to biofilm-associated infections in clinics and biofouling in industrial settings. This review focuses on describing biofilm adhesion strategies among different bacteria, including Vibrio cholerae, Pseudomonas aeruginosa, and Staphylococcus aureus. Techniques used to characterize biofilm adhesion are also reviewed. An understanding of biofilm adhesion strategies can guide the development of novel approaches to inhibit or manipulate biofilm adhesion and growth.

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