Axial filament involvement in the motility of Leptospira interrogans

Bromley, D B; Charon, Nyles W.

doi:10.1128/jb.137.3.1406-1412.1979

Cited by 69 publications

(28 citation statements)

References 15 publications

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“…However, the L. biflexa flaB mutants differ from the L. biflexa wild-type strain in that they are relatively straight, not flexible and without hook-shaped ends. Such linear mutants were also observed in Leptonema illini, belonging to the same Leptospiraceae family as Leptospira spp., after nitrosoguanidine treatment (Bromley and Charon, 1979). The characteristic hook-and spiral-shaped ends of Leptospira spp.…”

Section: Discussionmentioning

confidence: 99%

First evidence for gene replacement in Leptospira spp. Inactivation of L. biflexa flaB results in non‐motile mutants deficient in endoflagella

Picardeau¹,

Brenot

Girons

2001

Molecular Microbiology

138

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SummaryLeptospira spp. offer many advantages as model bacteria for the study of spirochaetes. However, homologous recombination between introduced DNA and the corresponding chromosomal loci has never been demonstrated. A unique feature of spirochaetes is the presence of endoflagella between the outer membrane sheath and the cell cylinder. We chose the flaB flagellin gene, constituting the flagellar core, as a target for gene inactivation in the saprophyte Leptospira biflexa. The amino acid sequence of the FlaB protein of L. biflexa was most similar to those of spirochaetes Brachyspira hyodysenteriae (agent of swine dysentery), Leptospira interrogans (agent of leptospirosis) and Treponema pallidum (agent of syphilis). A suicide vector containing the L. biflexa flaB gene disrupted by a kanamycin marker was UV irradiated or alkali denatured before electroporation. This methodology allowed the selection of many kanamycin-resistant colonies resulting from single and double cross-over events at the flaB locus. The double recombinant mutants are nonmotile, as visualized in both liquid and semi-solid media. In addition, a flaB mutant selected for further analysis was shown to be deficient in endoflagella by electron microscopy. However, most of the transformants had resulted from a single homologous recombination event, giving rise to the integration of the suicide vector. We evaluated the effect of the sacB and rpsL genes in L. biflexa as potential counterselectable markers for allelic exchange, and then used the rpsL system for the positive selection of flaB double recombinants in a streptomycinresistant strain. Like the flaB mutant studied above, the Str r double cross-over mutant was non-motile and deficient in endoflagella. Our results demonstrate that FlaB is involved in flagella assembly and motility. They also show the feasibility of performing allelic replacement in Leptospira spp. by homologous recombination.

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Section: Discussionmentioning

confidence: 99%

First evidence for gene replacement in Leptospira spp. Inactivation of L. biflexa flaB results in non‐motile mutants deficient in endoflagella

Picardeau¹,

Brenot

Girons

2001

Molecular Microbiology

138

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“…lc); a helix is right-handed if an object on it spirals clockwise (CW) when moving away from the observer [Carleton et al, 1979;Kayser and Adrian, 1978;Yoshii, 19781. Recent evidence suggests that the PFs do not overlap in the center of the cell [Birch-Andersen et al, 1973;Bromley and Charon, 1979;Hovind-Hougen, 19761. As with some other spirochetes [Bharier and Allis, 1974;Greenberg et al, 1985;Hardy et al, 1975;Joseph and Canale-Parola, 1972;Limberger and Charon, 1986;Penn et al, 1985;Radolf et al, 19861, but in contrast to most bacterial flagella [Doetsch and Sjoblad, 1980;Trachtenberg et al, 19861, multiple major protein species are associated with the PFs [Nauman et al, 19691.…”

Section: Leptospira Morphology and Motilitymentioning

confidence: 99%

Motility of the spirochete leptospira

Goldstein

Charon

1988

Cell Motil. Cytoskeleton

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Spirochetes are a group of bacteria with a unique ultrastructure and a fascinating swimming behavior. This article reviews the hydrodynamics of spirochete motility, and examines the motility of the spirochete Leptospira in detail. Models of Leprospiru motility are discussed, and future experiments are proposed.The outermost structure of Leptospiru is a membrane sheath, and within this sheath are a helically shaped cell cylinder and two periplasmic flagella. One periplasmic flagellum is attached subterminally at either end of the cell cylinder and extends partway down the length of the cell. In swimming cells, each end of the cell may assume either a spiral or a hook shape. Translational cells have the anterior end spiral shaped, and the posterior end hook shaped. In the model of Berg et al., the periplasmic flagella are believed to rotate between the sheath and the cell cylinder. Rotation of the anterior periplasmic flagellum causes the generation of a gyrating spiral-shaped wave. This wave is believed sufficient to propel the cells forward in a low-viscosity medium. The cell cylinder concomitantly rolls around the periplasmic flagella in the opposite direction-which allows the cell to literally screw through a gel-like viscous medium without slippage. This model is presented, and it is contrasted to previous models of Leptospiru motility.

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“…Spirochete PF are similar in structure and function to flagella of externally flagellated bacteria, as each consists of a basal body complex or motor, a flexible hook and a flagellar filament (Charon and Goldstein, 2002;Limberger, 2004). Whereas PF from non-Leptospira spirochetes display curved forms when viewed by electron microscopy once purified (Charon et al, 1991;Li et al, 2000a), leptospire PF are instead extensively supercoiled in the form of a spring (Berg et al, 1978;Bromley and Charon, 1979;Wolgemuth et al, 2006;Kan ad Wolgemuth, 2007). Furthermore, leptospire mutants that form uncoiled PF, or lacking PF altogether, maintain their helical cell body shape but display straight cell axes, and are also unable to generate translational motility (Bromley and Charon, 1979;Picardeau et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

A novel flagellar sheath protein, FcpA, determines filament coiling, translational motility and virulence for the Leptospira spirochete

Wunder

Figueira

Benaroudj

et al. 2016

Molecular Microbiology

120

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Summary Leptospira are unique among bacteria based on their helical cell morphology with hook-shaped ends and the presence of periplasmic flagella (PF) with pronounced spontaneous supercoiling. The factors that provoke such supercoiling, as well as the role that PF coiling plays in generating the characteristic hook-end cell morphology and motility, have not been elucidated. We have now identified an abundant protein from the pathogen L. interrogans, exposed on the PF surface, and named it Flagellar-coiling protein A (FcpA). The gene encoding FcpA is highly conserved among Leptospira and was not found in other bacteria. fcpA- mutants, obtained from clinical isolates or by allelic exchange, had relatively straight, smaller-diameter PF, and were not able to produce translational motility. These mutants lost their ability to cause disease in the standard hamster model of leptospirosis. Complementation of fcpA restored the wild-type morphology, motility and virulence phenotypes. In summary, we identified a novel Leptospira 36-kDa protein, the main component of the spirochete's PF sheath, and a key determinant of the flagella's coiled structure. FcpA is essential for bacterial translational motility and to enable the spirochete to penetrate the host, traverse tissue barriers, disseminate to cause systemic infection, and reach target organs.

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Axial filament involvement in the motility of Leptospira interrogans

Cited by 69 publications

References 15 publications

First evidence for gene replacement in Leptospira spp. Inactivation of L. biflexa flaB results in non‐motile mutants deficient in endoflagella

First evidence for gene replacement in Leptospira spp. Inactivation of L. biflexa flaB results in non‐motile mutants deficient in endoflagella

Motility of the spirochete leptospira

A novel flagellar sheath protein, FcpA, determines filament coiling, translational motility and virulence for the Leptospira spirochete

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