Flagella-dependent gliding motility inChlamydomonas

Summary Flagella are generally recognized as organelles of motility responsible for the ability ofChlamydomonas to swim through its environment. However, the same flagella are also responsible for an alternative form of whole cell locomotion, termed gliding. Use of paralyzed flagella mutants demonstrates that gliding is independent of axonemal bend propagation. Gliding motility results from an interaction of the flagellar surface with a solid substrate. Gliding is characterized by bidirectional movements at 1.6±0.3 m/second and occurs when the cell is in a characteristic gliding configuration, where the two flagella are oriented at 180° to one another. A variety of observations suggest that the leading flagellum is responsible for the force transduction resulting in cell locomotion, although both flagella have the capacity to function as the active flagellum. The characteristics of gliding motility have been compared with theChlamydomonas flagellar surface motility phenomenon defined as surface translocation of polystyrene microspheres.     

Substratum adhesion and gliding in a diatom are mediated by extracellular proteoglycans

Diatoms are unicellular microalgae encased in a siliceous cell wall, or frustule. Pennate diatoms, which possess bilateral symmetry, attach to the substratum at a slit in the frustule called the raphe. These diatoms not only adhere, but glide across surfaces whilst maintaining their attachment, secreting a sticky mucilage that forms a trail behind the gliding cells. We have raised monoclonal antibodies to the major cell surface proteoglycans of the marine raphid diatom Stauroneis decipiens Hustedt. The antibody StF.H4 binds to the cell surface, in the raphe and to adhesive trails and inhibits the ability of living diatoms to adhere to the substratum and to glide. Moreover, StF.H4 binds to a periodate-insensitive epitope on four frustule-associated proteoglycans (relative molecular masses 87, 112, and >200 kDa). Another monoclonal antibody, StF.D5, binds to a carbohydrate epitope on the same set of proteoglycans, although the antibody binds only to the outer surface of the frustule and does not inhibit cell motility and adhesion. Received: 2 December 1996 / Accepted: 6 March 1997     

Continuous observations of bacterial gliding motility in a dialysis microchamber: The effects of inhibitors

Use of a dialysis microchamber has allowed continuous observations on the same set of gliding bacteria during changes in the composition of the perfused medium. This procedure has revealed the presence of an adaptive, cyanide-insensitive metabolic pathway, which allows cyanide-treated Flexibacter BH3 to begin gliding again at a reduced rate when glucose is the substrate. In addition, it has revealed that individual flexibacter cells can maintain their gliding motility for up to 20 h in the absence of exogenous substrate.Gliding in Flexibacter BH3 was prevented by those inhibitors blocking the electron transport process. Inhibitors of glucose metabolism did not prevent motility, since the flexibacters obviously metabolize endogenous substrate under such circumstances. Proton ionophores, which induce membrane depolarization, rapidly inhibited gliding in Flexibacter BH3. This inhibition was irreversible in the case of gramicidin S. Gliding was not inhibited by cytochalasin B or antiactin antibody. High concentrations of Ca²⁺ were particularly inhibitory to the gliding process. The significance of these results is discussed in relation to a possible mechanism of gliding involving the generation of rhythmical contractions in the outer cell membrane of Flexibacter BH3.Abbreviations used CCCP carbonyl cyanide m-chlorophenyl hydrazone - DNP p-dinitrophenol - GMCS gramicidin S - HQNO 2-heptyl-4-hydroxyquinoline N-oxide - PCMB p-chloromercuribenzoate - CM complete Lewin's medium - BS Lewin's basal salts     

Inhibition of Cytophaga U67 gliding motility by inhibitors of polypeptide synthesis

Gliding motility of Cytophaga U67 and several other cytophagas was inhibited by a growth-permissive concentration of chloramphenicol (50 g/ml). Several other inhibitors of polypeptide synthesis also demonstrated this effect. Short-term exposure to several of these inhibitors resulted in reversible inhibition of gliding by growing cells. In wet mounts chloramphenicol-grown cells demonstrated non-translocational tumbling. Electrophoretic patterns of polypeptides released by ethylenediaminetetraacetate treatment of control and chloramphenicol-grown cells were distinct. Gliding of a spontaneous mutant was resistant to chloramphenicol at 50 g/ml; its motility was inhibited at the growth-permissive concentration of 400 g/ml.     

Yaw stability in gliding birds

A new concept for describing the yaw stability in gliding birds is presented. This concept introduces dynamic stiffness in yaw as an appropriate indication of stability. Other than the conventional metric of static yaw stability given by the gradient of the aerodynamic yawing moment with respect to the sideslip angle, the dynamic stiffness does not only provide a qualitative indication of stability but also a precise quantitative measure of the restoring action in the yaw axis. With the use of scaling relations, it is shown that the dynamic stiffness of birds is sufficiently high though their static yaw stability may be very small. The underlying mechanism is that the yaw moment of inertia is more reduced with a decrease in size than the restoring aerodynamic moment. Reference is made to the yaw stability in aircraft and related flying qualities requirements. Thus, numerical values are derived which can be used as a standard of comparison providing a rating basis for the dynamic yaw stiffness in small flying objects, like birds. Furthermore, it is shown that the wings of birds produce yawing moments due to sideslip so large that a sufficiently high level of dynamic yaw stiffness can be achieved. From the results derived in this paper, it may be concluded that birds—unlike aircraft—need no vertical tail for yaw stability.     

Structural specificity of sugars that inhibit gliding motility of Cytophaga johnsonae

It is a common observation that gliding bacteria form raised, smooth-edged colonies on nutrient-rich media, and typical thin, spreading, uneven-edged colonies on nutrient-poor media. An earlier study of the effect of different sugars on colony spreading by Cytophaga johnsonae was expanded to include the effects of several sugars and other organic compounds on the motility of groups of cells (rafts), and latex bead movement on cells' surfaces. When the structures of those sugars that did, or did not, affect raft formation and colony spreading were compared, it was noted that those sugars that inhibited these two manifestations of gliding motility all possessed a common sub-structure, that found in the portion of glucopyranose comprising carbons 3, 4, 5, and 6. If these structural features were altered chemically or stereochemically, the resulting molecule had little to no effect on motility. The differential effects of some compounds on raft formation, colony spreading, and bead movement are noted. A regulatory mechanism that would turn off motility in the presence of an inhibitory sugar is implicated, and the relevance of such a system to the life of the organism is discussed. We report, as well, additional compounds that will serve as carbon and energy sources for C. johnsonae.     

Biochemical characterization of a mutant of the yeast Pichia anomala derepressed for malic acid utilization in the presence of glucose

Abstract Myxococcus xanthus cells move over surfaces by gliding motility. The frz signal transduction system is used to control the reversal frequency, and thus the overall direction of movement of M. xanthus cells. We analyzed the behavior of wild-type and frz mutant cells in response to prey bacteria ( Escherichia coli ). Wild-type cells of M. xanthus did not respond to microcolonies of E. coli until they made physical contact. Cells which penetrated a colony remained in the colony until all of the prey cells were digested. Cells of frz mutants also penetrated E. coli microcolonies and digested some of the E. coli cells, but they invariably abandoned the microcolony leaving their food source behind. These observations illustrate the importance of the frz system of signal transduction for the feeding behavior of M. xanthus cells.     

Genetics of gliding motility in Myxococcus xanthus: Molecular cloning of the mgl locus

Summary Wild-type Myxococcus xanthus cells move across solid surfaces by gliding. However no locomotory organelles for gliding have as yet been identified. Two sets of genes are required for gliding in M. xanthus: Gene System A is necessary for the gliding of isolated cells and Gene System S comes into play when cells are close together. The product of the mgl locus is required for both types of gliding and therefore may be a structural component of the gliding organelle. To begin to investigate the function of mgl in gliding a 12 kb segment of M. xanthus DNA containing the locus was cloned in Escherichia coli and returned to Myxococcus by specialized transduction with coliphage P1. In M. xanthus the chimeric plasmid integrates into the chromosome by recombination between the cloned segment and its homolog in the recipient chromosome forming a tandem duplication of the cloned segment with the vector sequences at the novel joint. The construction of partial diploids in this manner facilitated dominance tests and interallelic crosses with ten mgl alleles. We also describe a method for the analysis of tandem duplications that precisely maps alleles to a specific copy of the duplicated sequences. This method provides evidence for the dominance of mgl ⁺ over the mgl ^- alleles. It also reveals what appears to be gene conversion at this locus during recombination between a cloned mgl sequence and its homolog in the chromosome.     

Purification of the N-acylaminosulfonates,unusual lipids of gliding bacteria

A group of unusual sulfonolipids that are major components of the cell envelope has been found in gliding bacteria of the genus Cytophaga and closely related genera. One of the lipids, capnine, has been previously isolated and shown to be 2-amino-3-hydroxy-15-methylhexadecane-1-sulfonic acid. Capnine accumulates in large quantities in some Capnocytophaga spp., but in other organisms the predominant sulfonolipids are compounds that are less polar than capnine and lack the free amino group of that compound. A method is described for the purification of these less polar lipids employing chromatography on hydroxylapatite and DEAE-cellulose in organic solvents. The sulfonolipids have been isolated in high yield and essentially pure from two Cytophaga spp., one Flexibacter sp. and one Capnocytophaga sp. Preliminary characterization of the lipids of infrared absorption spectroscopy, thin layer chromatography and other methods has shown them to be N-acylaminosulfonates, the aminosulfonate moiety of which is closely related (if not identical) to capnine.     

Calcium requirement for gliding motility in myxobacteria.

The ability to glide on a solid surface was inducible by calcium ion in Stigmatella aurantiaca. The induction of motility but not motility itself was prevented by chloramphenicol and erythromycin. Calcium ion was also required for cells to glide, even when they were previously induced. The ability of Myxococcus xanthus to glide in groups using the S motility system but not as single cells (A system) was prevented by chloramphenicol and erythromycin.     

A conjecture on the relationship of bacterial shape to motility in rod-shaped bacteria

We have calculated the optimal shape, i.e. the length-to-width ratio of a bacterial cell, that allows a bacterial cell to move most efficiently through liquid. For a cell of a given size, a minimum exists in the force required to move through any liquid when the length of the cell is approx. 3.7 times greater than the width. As this is in approximate agreement with the observed shape of bacteria such as the Enterobacteriaceae, we conjecture that the current observed shape of these bacteria may have been determined, in part, to obtain the most efficient shape for moving through liquids. It is also found that spherical cells are very inefficient in movement through liquid, while longer cells of a fixed size are still relatively efficient in moving through liquids. Since the optimal shape is independent of actual size (within large bounds), it is further proposed that hydrodynamic efficiency considerations support the proposal of constant shape over a range of sizes for rod-shaped bacteria.     

Ghrelin as a target for gastrointestinal motility disorders

The therapeutic potential of ghrelin and synthetic ghrelin receptor (GRLN-R) agonists for the treatment of gastrointestinal (GI) motility disorders is based on their ability to stimulate coordinated patterns of propulsive GI motility. This review focuses on the latest findings that support the therapeutic potential of GRLN-R agonists for the treatment of GI motility disorders. The review highlights the preclinical and clinical prokinetic effects of ghrelin and a series of novel ghrelin mimetics to exert prokinetic effects on the GI tract. We build upon a series of excellent reviews to critically discuss the evidence that supports the potential of GRLN-R agonists to normalize GI motility in patients with GI hypomotility disorders such as gastroparesis, post-operative ileus (POI), idiopathic chronic constipation and functional bowel disorders.     

Nutritional studies on Chloroflexus,a filamentous photosynthetic,gliding bacterium

Nutritional studies on four different strains of Chloroflexus, a new genus of filamentous, photosynthetic bacteria are described. This organism appears to be related to several different procaryotic groups, and in particular to the green sulfur bacteria and blue-green algae. Unlike these autotrophs, however, Chloroflexus is nutritionally diverse, being able to grow aerobically as a light-independent heterotroph, and anaerobically as a photoautotroph or photoheterotroph. Numerous organic carbon sources including hexoses, amino acids, short chain fatty acids, organic acids, and some alcohols are utilized under various growth conditions. These results suggest that this organism may be among the most nutritionally versatile organisms known.     

Effects of solar and ultraviolet radiation on motility, photomovement and pigmentation in filamentous, gliding cyanobacteria

Abstract The effects of solar irradiation and artificial UV irradiation on several cyanobacteria ( Anabaena variabilis and two strains of Phormidium uncinatum ) have been studied. Both types of radiation affect the percentage of motile filaments and impair the linear velocity of the organisms. Long term exposure to UV radiation bleaches the photosynthetic pigments as determined by absorption difference spectra. Fluorescence excitation and emission spectra indicate that under ultraviolet radiation the energy transfer from the accessory pigments to chlorophyll is affected. Furthermore the structural integrity of the phycobilisomes seems to be impaired by continuous radiation and the photoreceptor pigments seem to be destroyed.     

Extreme UV-A sensitivity of the filamentous gliding bacterium Vitreoscilla stercoraria

Strains of the filamentous gliding bacterium Vitreoscilla, LB13 and C1, are shown to be highly sensitive to UV-A (320-400 nm), with an LD50 of less than 20 kJ m(-2). Vitreoscilla LB13 can be protected from UV-A by including superoxide dismutase and catalase, separately or in combination, during the exposure, indicating an involvement of reactive oxygen species. LB13A, a photo-insensitive strain derived from LB13, is described.     

Cell motility assays

This report summarises practical aspects to measuring cell motility in culture. The methods described here were discussed at a 1-day European Tissue Culture Society (ETCS-UK) workshop organised by John Masters and Gareth E Jones that was held at University College London on 19th April 2007.     

Protein content of peptidoglycan of liquid-grown cells differs from that of surface-grown,gliding Cytophaga johnsonae

The peptidoglycan sacculi of surface-grown Cytophaga johnsonae had associated with them a large amoutn of protein (the major species is 50 kDa) whereas sacculi from liquid-grown cells had little or no attached protein. The 50 kDa protein was localized in the outer membrane of liquid-grown cells. A portion of this membrane-derived 50 kDa protein was attached to the peptidoglycan only when the cells made contact with the substratum. Protein synthesis did not appear to be required for attachment as the process was not inhibited by chloramphenicol. Association of the 50 kDa protein with the peptidoglycan in response to cell contact with the substratum is suggested.