scholarly journals Biofilm Spreading by the Adhesin-Dependent Gliding Motility of Flavobacterium johnsoniae. 1. Internal Structure of the Biofilm

2021 ◽  
Vol 22 (4) ◽  
pp. 1894
Author(s):  
Keiko Sato ◽  
Masami Naya ◽  
Yuri Hatano ◽  
Yoshio Kondo ◽  
Mari Sato ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Leading Edge ◽  
Time Lapse ◽  
Gliding Motility ◽  
Solid Surfaces ◽  
Negative Bacterium ◽  
Wild Type ◽  
Thick Fiber ◽  
Flavobacterium Johnsoniae ◽  
Vesicle Secretion

The Gram-negative bacterium Flavobacterium johnsoniae employs gliding motility to move rapidly over solid surfaces. Gliding involves the movement of the adhesin SprB along the cell surface. F. johnsoniae spreads on nutrient-poor 1% agar-PY2, forming a thin film-like colony. We used electron microscopy and time-lapse fluorescence microscopy to investigate the structure of colonies formed by wild-type (WT) F. johnsoniae and by the sprB mutant (ΔsprB). In both cases, the bacteria were buried in the extracellular polymeric matrix (EPM) covering the top of the colony. In the spreading WT colonies, the EPM included a thick fiber framework and vesicles, revealing the formation of a biofilm, which is probably required for the spreading movement. Specific paths that were followed by bacterial clusters were observed at the leading edge of colonies, and abundant vesicle secretion and subsequent matrix formation were suggested. EPM-free channels were formed in upward biofilm protrusions, probably for cell migration. In the nonspreading ΔsprB colonies, cells were tightly packed in layers and the intercellular space was occupied by less matrix, indicating immature biofilm. This result suggests that SprB is not necessary for biofilm formation. We conclude that F. johnsoniae cells use gliding motility to spread and maturate biofilms.

scholarly journals The Type 9 Secretion System Is Required for Flavobacterium johnsoniae Biofilm Formation

2021 ◽  
Vol 12 ◽  
Author(s):  
Todd J. Eckroat ◽  
Camillus Greguske ◽  
David W. Hunnicutt
Keyword(s):  
Biofilm Formation ◽  
Deletion Mutant ◽  
Secretion System ◽  
Gliding Motility ◽  
Microplate Assay ◽  
Partial Reduction ◽  
Wild Type ◽  
Flavobacterium Johnsoniae ◽  
Biofilm Production ◽  
Type Ix Secretion System

Flavobacterium johnsoniae forms biofilms in low nutrient conditions. Protein secretion and cell motility may have roles in biofilm formation. The F. johnsoniae type IX secretion system (T9SS) is important for both secretion and motility. To determine the roles of each process in biofilm formation, mutants defective in secretion, in motility, or in both processes were tested for their effects on biofilm production using a crystal violet microplate assay. All mutants that lacked both motility and T9SS-mediated secretion failed to produce biofilms. A porV deletion mutant, which was severely defective for secretion, but was competent for motility, also produced negligible biofilm. In contrast, mutants that retained secretion but had defects in gliding formed biofilms. An sprB mutant that is severely but incompletely defective in gliding motility but retains a fully functional T9SS was similar to the wild type in biofilm formation. Mutants with truncations of the gldJ gene that compromise motility but not secretion showed partial reduction in biofilm formation compared to wild type. Unlike the sprB mutant, these gldJ truncation mutants were essentially nonmotile. The results show that a functional T9SS is required for biofilm formation. Gliding motility, while not required for biofilm formation, also appears to contribute to formation of a robust biofilm.

scholarly journals Colony spreading of the gliding bacterium Flavobacterium johnsoniae in the absence of the motility adhesin SprB

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Keiko Sato ◽  
Masami Naya ◽  
Yuri Hatano ◽  
Yoshio Kondo ◽  
Mari Sato ◽  
...  
Keyword(s):  
Soft Agar ◽  
Gliding Motility ◽  
Wild Type ◽  
Initial Cell ◽  
Flavobacterium Johnsoniae ◽  
Seeking Behavior ◽  
Colony Spreading ◽  
Gliding Bacterium ◽  
Mutant Cells ◽  
Scanning Electron

AbstractColony spreading of Flavobacterium johnsoniae is shown to include gliding motility using the cell surface adhesin SprB, and is drastically affected by agar and glucose concentrations. Wild-type (WT) and ΔsprB mutant cells formed nonspreading colonies on soft agar, but spreading dendritic colonies on soft agar containing glucose. In the presence of glucose, an initial cell growth-dependent phase was followed by a secondary SprB-independent, gliding motility-dependent phase. The branching pattern of a ΔsprB colony was less complex than the pattern formed by the WT. Mesoscopic and microstructural information was obtained by atmospheric scanning electron microscopy (ASEM) and transmission EM, respectively. In the growth-dependent phase of WT colonies, dendritic tips spread rapidly by the movement of individual cells. In the following SprB-independent phase, leading tips were extended outwards by the movement of dynamic windmill-like rolling centers, and the lipoproteins were expressed more abundantly. Dark spots in WT cells during the growth-dependent spreading phase were not observed in the SprB-independent phase. Various mutations showed that the lipoproteins and the motility machinery were necessary for SprB-independent spreading. Overall, SprB-independent colony spreading is influenced by the lipoproteins, some of which are involved in the gliding machinery, and medium conditions, which together determine the nutrient-seeking behavior.

scholarly journals Cloning and Characterization of theFlavobacterium johnsoniae Gliding Motility GenesgldD and gldE

2001 ◽  
Vol 183 (14) ◽  
pp. 4167-4175 ◽  
Author(s):  
David W. Hunnicutt ◽  
Mark J. McBride
Keyword(s):  
Membrane Protein ◽  
Cytoplasmic Membrane ◽  
Sequence Similarity ◽  
Gliding Motility ◽  
Wild Type ◽  
Bacteriophage Infection ◽  
Flavobacterium Johnsoniae ◽  
Latex Spheres ◽  
Serovar Typhimurium

ABSTRACT Cells of Flavobacterium johnsoniae move over surfaces by a process known as gliding motility. The mechanism of this form of motility is not known. Cells of F. johnsoniaepropel latex spheres along their surfaces, which is thought to be a manifestation of the motility machinery. Three of the genes that are required for F. johnsoniae gliding motility,gldA, gldB, and ftsX, have recently been described. Tn4351 mutagenesis was used to identify another gene, gldD, that is needed for gliding. Tn4351-induced gldD mutants formed nonspreading colonies, and cells failed to glide. They also lacked the ability to propel latex spheres and were resistant to bacteriophages that infect wild-type cells. Introduction of wild-type gldD into the mutants restored motility, ability to propel latex spheres, and sensitivity to bacteriophage infection. gldD codes for a cytoplasmic membrane protein that does not exhibit strong sequence similarity to proteins of known function. gldE, which lies immediately upstream ofgldD, encodes another cytoplasmic membrane protein that may be involved in gliding motility. Overexpression ofgldE partially suppressed the motility defects of agldB point mutant, suggesting that GldB and GldE may interact. GldE exhibits sequence similarity to Borrelia burgdorferi TlyC and Salmonella enterica serovar Typhimurium CorC.

scholarly journals Flavobacterium johnsoniae Gliding Motility Genes Identified by mariner Mutagenesis

2005 ◽  
Vol 187 (20) ◽  
pp. 6943-6952 ◽  
Author(s):  
Timothy F. Braun ◽  
Manjeet K. Khubbar ◽  
Daad A. Saffarini ◽  
Mark J. McBride
Keyword(s):  
Complete Loss ◽  
Gliding Motility ◽  
Wild Type ◽  
Flavobacterium Johnsoniae ◽  
Chemically Induced

ABSTRACT Cells of Flavobacterium johnsoniae glide rapidly over surfaces. The mechanism of F. johnsoniae gliding motility is not known. Eight gld genes required for gliding motility have been described. Disruption of any of these genes results in complete loss of gliding motility, deficiency in chitin utilization, and resistance to bacteriophages that infect wild-type cells. Two modified mariner transposons, HimarEm1 and HimarEm2, were constructed to allow the identification of additional motility genes. HimarEm1 and HimarEm2 each transposed in F. johnsoniae, and nonmotile mutants were identified and analyzed. Four novel motility genes, gldK, gldL, gldM, and gldN, were identified. GldK is similar in sequence to the lipoprotein GldJ, which is required for gliding. GldL, GldM, and GldN are not similar in sequence to proteins of known function. Cells with mutations in gldK, gldL, gldM, and gldN were defective in motility and chitin utilization and were resistant to bacteriophages that infect wild-type cells. Introduction of gldA, gldB, gldD, gldFG, gldH, gldI, and gldJ and the region spanning gldK, gldL, gldM, and gldN individually into 50 spontaneous and chemically induced nonmotile mutants restored motility to each of them, suggesting that few additional F. johnsoniae gld genes remain to be identified.

scholarly journals Gliding Motility Revisited: How Do the Myxobacteria Move without Flagella?

2010 ◽  
Vol 74 (2) ◽  
pp. 229-249 ◽  
Author(s):  
Emilia M. F. Mauriello ◽  
Tâm Mignot ◽  
Zhaomin Yang ◽  
David R. Zusman
Keyword(s):  
Biofilm Formation ◽  
Protein Localization ◽  
Myxococcus Xanthus ◽  
Gliding Motility ◽  
Solid Surfaces ◽  
Biological Functions ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Periplasmic Flagella ◽  
Motile Bacterium

SUMMARY In bacteria, motility is important for a wide variety of biological functions such as virulence, fruiting body formation, and biofilm formation. While most bacteria move by using specialized appendages, usually external or periplasmic flagella, some bacteria use other mechanisms for their movements that are less well characterized. These mechanisms do not always exhibit obvious motility structures. Myxococcus xanthus is a motile bacterium that does not produce flagella but glides slowly over solid surfaces. How M. xanthus moves has remained a puzzle that has challenged microbiologists for over 50 years. Fortunately, recent advances in the analysis of motility mutants, bioinformatics, and protein localization have revealed likely mechanisms for the two M. xanthus motility systems. These results are summarized in this review.

scholarly journals Flavobacterium johnsoniae gldN and gldO Are Partially Redundant Genes Required for Gliding Motility and Surface Localization of SprB

2009 ◽  
Vol 192 (5) ◽  
pp. 1201-1211 ◽  
Author(s):  
Ryan G. Rhodes ◽  
Mudiarasan Napoleon Samarasam ◽  
Abhishek Shrivastava ◽  
Jessica M. van Baaren ◽  
Soumya Pochiraju ◽  
...  
Keyword(s):  
Protein Secretion ◽  
Deletion Mutant ◽  
Gliding Motility ◽  
Selection Strategy ◽  
Wild Type ◽  
Flavobacterium Johnsoniae ◽  
Redundant Genes ◽  
Surface Localization ◽  
Gliding Bacterium ◽  
Mutant Cells

ABSTRACT Cells of the gliding bacterium Flavobacterium johnsoniae move rapidly over surfaces. Mutations in gldN cause a partial defect in gliding. A novel bacteriophage selection strategy was used to aid construction of a strain with a deletion spanning gldN and the closely related gene gldO in an otherwise wild-type F. johnsoniae UW101 background. Bacteriophage transduction was used to move a gldN mutation into F. johnsoniae UW101 to allow phenotypic comparison with the gldNO deletion mutant. Cells of the gldN mutant formed nonspreading colonies on agar but retained some ability to glide in wet mounts. In contrast, cells of the gldNO deletion mutant were completely nonmotile, indicating that cells require GldN, or the GldN-like protein GldO, to glide. Recent results suggest that Porphyromonas gingivalis PorN, which is similar in sequence to GldN, has a role in protein secretion across the outer membrane. Cells of the F. johnsoniae gldNO deletion mutant were defective in localization of the motility protein SprB to the cell surface, suggesting that GldN may be involved in secretion of components of the motility machinery. Cells of the gldNO deletion mutant were also deficient in chitin utilization and were resistant to infection by bacteriophages, phenotypes that may also be related to defects in protein secretion.

scholarly journals The dgc2 gene encoding di-guanylate cyclase suppresses both motility and biofilm formation in the filamentous cyanobacterium Leptolyngbya boryana.

2022 ◽  
Author(s):  
Kazuma Toida ◽  
Wakana Kushida ◽  
Hiroki Yamamoto ◽  
Kyoka Yamamoto ◽  
Kazuma Uesaka ◽  
...  
Keyword(s):  
Mutant Strain ◽  
Biofilm Formation ◽  
Catalytic Domain ◽  
Genetic Model ◽  
Model Organism ◽  
Gliding Motility ◽  
Spontaneous Mutant ◽  
Filamentous Cyanobacterium ◽  
Wild Type

Colony pattern formations of bacteria with motility manifest complicated morphological self-organization phenomena. Leptolyngbya boryana is the filamentous cyanobacterial species, which has been used as a genetic model organism for studying metabolism including photosynthesis and nitrogen-fixation. Although a widely used type strain (wild type) of this species has not been reported to show any motile activity, we isolated a spontaneous mutant strain which shows active motility (gliding activity) to give rise to complicated colony patters, including comet-like wandering clusters and disk-like rotating vortices on solid media. Whole-genome resequencing identified multiple mutations on the genome in the mutant strain. We confirmed that inactivation of a candidate gene, dgc2 (LBDG_02920), in the wild type background was sufficient to give rise to motility and the morphological colony patterns. This gene encodes a protein, containing the GGDEF motif, which is conserved at the catalytic domain of diguanylate cyclase (DGC). Although DGC has been reported to be involved in biofilm formation, the mutant strain lacking dgc2 significantly facilitated biofilm formation, suggesting a role of DGC for suppressing both gliding motility and biofilm formation. Thus, L. boryana provides an excellent genetic model to study dynamic colony pattern formation, and novel insight on a role of c-di-GMP for biofilm formation.

scholarly journals Social motility of biofilm-like microcolonies in a gliding bacterium

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chao Li ◽  
Amanda Hurley ◽  
Wei Hu ◽  
Jay W. Warrick ◽  
Gabriel L. Lozano ◽  
...  
Keyword(s):  
Self Assembly ◽  
Microscopic Analysis ◽  
Extracellular Polysaccharide ◽  
Microfluidic System ◽  
Gliding Motility ◽  
Solid Surfaces ◽  
Swarming Motility ◽  
Collective Movement ◽  
Flavobacterium Johnsoniae ◽  
Gliding Bacterium

AbstractBacterial biofilms are aggregates of surface-associated cells embedded in an extracellular polysaccharide (EPS) matrix, and are typically stationary. Studies of bacterial collective movement have largely focused on swarming motility mediated by flagella or pili, in the absence of a biofilm. Here, we describe a unique mode of collective movement by a self-propelled, surface-associated biofilm-like multicellular structure. Flavobacterium johnsoniae cells, which move by gliding motility, self-assemble into spherical microcolonies with EPS cores when observed by an under-oil open microfluidic system. Small microcolonies merge, creating larger ones. Microscopic analysis and computer simulation indicate that microcolonies move by cells at the base of the structure, attached to the surface by one pole of the cell. Biochemical and mutant analyses show that an active process drives microcolony self-assembly and motility, which depend on the bacterial gliding apparatus. We hypothesize that this mode of collective bacterial movement on solid surfaces may play potential roles in biofilm dynamics, bacterial cargo transport, or microbial adaptation. However, whether this collective motility occurs on plant roots or soil particles, the native environment for F. johnsoniae, is unknown.

scholarly journals SprB Is a Cell Surface Component of the Flavobacterium johnsoniae Gliding Motility Machinery

2008 ◽  
Vol 190 (8) ◽  
pp. 2851-2857 ◽  
Author(s):  
Shawn S. Nelson ◽  
Sreelekha Bollampalli ◽  
Mark J. McBride
Keyword(s):  
Cell Surface ◽  
Surface Protein ◽  
Gliding Motility ◽  
Wild Type ◽  
Polystyrene Microspheres ◽  
Flavobacterium Johnsoniae ◽  
A Cell ◽  
Transposon Insertions ◽  
Gliding Bacterium ◽  
Surface Adhesins

ABSTRACT Cells of the gliding bacterium Flavobacterium johnsoniae move rapidly over surfaces by an unknown mechanism. Transposon insertions in sprB resulted in cells that were defective in gliding. SprB is a highly repetitive 669-kDa cell surface protein, and antibodies against SprB inhibited the motility of wild-type cells. Polystyrene microspheres coated with antibodies against SprB attached to and were rapidly propelled along the cell surface, suggesting that SprB is one of the outermost components of the motility machinery. The movement of SprB along the cell surface supports a model of gliding motility in which motors anchored to the cell wall rapidly propel cell surface adhesins.

scholarly journals Involvement of the Type IX Secretion System in Capnocytophaga ochracea Gliding Motility and Biofilm Formation

2016 ◽  
Vol 82 (6) ◽  
pp. 1756-1766 ◽  
Author(s):  
Daichi Kita ◽  
Satoshi Shibata ◽  
Yuichiro Kikuchi ◽  
Eitoyo Kokubu ◽  
Koji Nakayama ◽  
...  
Keyword(s):  
Cell Surface ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Secretion System ◽  
Gliding Motility ◽  
Confocal Laser ◽  
Bacterial Cells ◽  
Wild Type ◽  
Content Type ◽  
Type Ix Secretion System

ABSTRACTCapnocytophaga ochraceais a Gram-negative, rod-shaped bacterium that demonstrates gliding motility when cultured on solid agar surfaces.C. ochraceapossesses the ability to form biofilms; however, factors involved in biofilm formation by this bacterium are unclear. A type IX secretion system (T9SS) inFlavobacterium johnsoniaewas shown to be involved in the transport of proteins (e.g., several adhesins) to the cell surface. Genes orthologous to those encoding T9SS proteins inF. johnsoniaehave been identified in the genome ofC. ochracea; therefore, the T9SS may be involved in biofilm formation byC. ochracea. Here we constructed three ortholog-deficientC. ochraceamutants lackingsprB(which encodes a gliding motility adhesin) orgldKorsprT(which encode T9SS proteins inF. johnsoniae). Gliding motility was lost in each mutant, suggesting that, inC. ochracea, the proteins encoded bysprB,gldK, andsprTare necessary for gliding motility, and SprB is transported to the cell surface by the T9SS. For the ΔgldK, ΔsprT, and ΔsprBstrains, the amounts of crystal violet-associated biofilm, relative to wild-type values, were 49%, 34%, and 65%, respectively, at 48 h. Confocal laser scanning and scanning electron microscopy revealed that the biofilms formed by wild-typeC. ochraceawere denser and bacterial cells were closer together than in those formed by the mutant strains. Together, these results indicate that proteins exported by the T9SS are key elements of the gliding motility and biofilm formation ofC. ochracea.

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