Effect of mechanical removal of pili on gliding motility of Myxococcus xanthus

Rosenbluh, A; Eisenbach, Michael

doi:10.1128/jb.174.16.5406-5413.1992

Cited by 25 publications

(24 citation statements)

References 31 publications

(24 reference statements)

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“…Pili were intentionally sheared off DK10469 cells by passage through a needle 50 times as previously described (39), except that a 25-gauge 7.5-cm spinal needle was used. Effectiveness of shearing was confirmed by electron microscopy, which showed a reduction in piliated cell ends to 1 to 3% (versus 30 to 40% for unsheared cells).…”

Section: Methodsmentioning

confidence: 99%

Regulation of expression of the pilA gene in Myxococcus xanthus

1997

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Type IV pili are required for social gliding motility in Myxococcus xanthus. In this work, the expression of pilin (the pilA gene product) during vegetative growth and fruiting-body development was examined. A polyclonal antibody against the pilA gene product (prepilin) was prepared, along with a pilA-lacZ fusion, and was used to assay expression of pilA in M. xanthus in different mutant backgrounds. pilA expression required the response regulator pilR but was negatively regulated by the putative sensor kinase pilS. pilA expression did not require pilB, pilC, or pilT. pilA was also autoregulated; a mutation which altered an invariant glutamate five residues from the presumed prepilin processing site eliminated this autoregulation, as did a deletion of the pilA gene. Primer extension and S1 nuclease analysis identified a 54 promoter upstream of pilA, consistent with the homology of pilR to the NtrC family of response regulators. Expression of pilA was found to be developmentally regulated; however, the timing of this expression pattern was not entirely dependent on pilS or pilR. Finally, pilA expression was induced by high nutrient concentrations, an effect that was also not dependent on pilS or pilR.

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Section: Methodsmentioning

confidence: 99%

Regulation of expression of the pilA gene in Myxococcus xanthus

1997

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“…Kaiser further described fimbriae as 10-nmlong polar appendages that were associated with social motility (22). Rosenbluh and Eisenbach confirmed that, while not required for the maintenance of cell-cell cohesions, pili appear to be involved in the coordination of the movement of cells translocating as groups (social motility) (35). Unlike fibrils (see below), fimbriae are expressed constitutively on cells grown at low densities and in liquid culture (35).…”

mentioning

confidence: 97%

Integral proteins of the extracellular matrix fibrils of Myxococcus xanthus

Behmlander

Dworkin

1994

J Bacteriol

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“…No mutant lacking pili retains social motility (21,56). Social motility is also lost when pili are detached by violent shearing and reappears when pili are allowed to grow back (41). Several social motility genes, including tgl (transient glider), the subject of this paper, are needed for production of pili.…”

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confidence: 99%

Identification and localization of the Tgl protein, which is required for Myxococcus xanthus social motility

Rodriguez-Soto

Kaiser

1997

J Bacteriol

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Tgl protein is required for the production of the type IV pili found at a pole of the Myxococcus xanthus cell. These pili are essential for social motility. Evidence is presented that Tgl is a membrane protein, based on experiments with polyclonal antibody specific for Tgl that was raised against the fusion proteins ␤-galactosidase-Tgl and TrpE-Tgl. Immunoaffinity-purified antibody reacted with a protein in M. xanthus having an apparent molecular mass of 27.5 kDa as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, while the sequence of the tgl gene translates into a polypeptide of 27 kDa. Although these numbers are close, it is likely that the primary tgl translation product is processed and modified in M. xanthus. The N terminus has a signal peptidase II recognition sequence, cleavage of which is expected to remove 19 amino acid residues. When the tgl gene is expressed in Escherichia coli, the protein product consistently migrates faster in the gel than mature Tgl expressed in M. xanthus, suggesting a second modification by addition which slows migration of the protein from M. xanthus. Tgl, as detected by its specific antibody, sediments with the membrane fraction of cells. It can be extracted with detergents but not with salt or by the addition of chelators for divalent cations. In an equilibrium gradient, Tgl bands at the buoyant density of membranes and with the NADH-oxidase activity. Intact cells failed to bind anti-Tgl antibody, and less than 2% of the total Tgl is released in soluble form from the periplasm. Yet, cells that had been osmotically shocked and treated with paraformaldehyde were able to react with the specific antibody-a reaction absent from cells with a deletion of the tgl transcription unit. Assuming that osmotic shock disrupts the outer membrane, the fractionation and localization data imply that Tgl is attached to the inner or outer membranes, from which it is exposed to the intermembranous space. Tgl is necessary for synthesis of pili in M. xanthus and is the only pilus protein that can be donated by other cells (stimulation). Tgl contains six tandem copies of the tetratrico peptide repeat structural motif. Its membrane localization, capacity for stimulation, and content of tetratrico structural repeats together suggest that Tgl may be necessary for the assembly of pilin subunits into filaments.

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Effect of mechanical removal of pili on gliding motility of Myxococcus xanthus

Cited by 25 publications

References 31 publications

Regulation of expression of the pilA gene in Myxococcus xanthus

Regulation of expression of the pilA gene in Myxococcus xanthus

Integral proteins of the extracellular matrix fibrils of Myxococcus xanthus

Identification and localization of the Tgl protein, which is required for Myxococcus xanthus social motility

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