Biochemistry of Bacterial Flagella

Dimmitt, K.; Emerson, Suzanne U.; Tokuyasu, K. T.

doi:10.1007/978-1-4684-1962-7_14

Cited by 2 publications

(2 citation statements)

References 23 publications

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“…However, very little is known about the biochemistry involved in these processes. The flagellar organelle can be isolated from Escherichia coli and Bacillus subtilis (7,8). It is composed of a long helical filament and a complex basal structure which is tightly bound to the outer membrane of the cell.…”

Section: Introductionmentioning

confidence: 99%

The identification of the mot gene product with Escherichia coli-lambda hybrids.

Silverman¹,

Matsumura²

1976

Proc. Natl. Acad. Sci. U.S.A.

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Molecular cloning techniques were used to construct lambda-E. coli hybrid bacteriophage carrying genes involved in bacterial flagellar motility (mot) and chemotaxis (cheA). A series of hybrid bacteriophage without each of these genes was also prepared. When paralyzed mutants of E. coli were infected with lambda that carried the mot gene, the ability of the bacterium to swim was rapidly restored. The restoration of motility was the result of the synthesis and insertion into the cell membrane of a protein with an apparent molecular weight of 31,000 (the Mot protein).Another polypeptide with a mobility on acrylamide gel electrophoresis which corresponded to a molecular weight of 39,000 was associated with the cheA gene. The presence of this polypeptide alone was not sufficient to restore chemotactic activity to mutant cheA strains. It was suggested that only a portion of the cheA gene was cloned, and thus the 39,000 protein may be a partial product of the cheA gene, or the product of a second mot gene.Bacterial flagella function by rotating, and their rotation is regulated so that it responds to information provided by specific chemoreceptors on the surface of the cell (1-3). The genetic basis for flagellar structure, function, and regulation has been analyzed in detail (4-6). However, very little is known about the biochemistry involved in these processes. The flagellar organelle can be isolated from Escherichia coli and Bacillus subtilis (7,8). It is composed of a long helical filament and a complex basal structure which is tightly bound to the outer membrane of the cell. The basal structure has been purified (9) and its components were characterized by gel electrophoresis. It was found to contain eleven different polypeptide chains (10). It is clear, however, that these components do not account for all of the gene products involved in flagellar function. This conclusion is based on a comparison of the proteins obtained from flagella isolated from mot amber mutants and wild-type strains. The mot gene appears to control the synthesis of a product which is directly involved in flagellar activity. Mutations in (16,17).To understand the mechanisms involved in motility, and in the assembly and function of flagella, it is necessary to identify the products of all of the genes that contribute to these processes.One approach involves an isolation of the genes and detection of their products directly. In a previous report (18), we showed that hybrid E. coli-lambda phage carrying genes that control flagellin synthesis could be obtained and used to determine the regulation of flagellin production. We will now show how hybrid lambda bacteriophage can be used to assay for other flagella-specific proteins which include the mot gene product. MATERIALS AND METHODSConstruction and Propagation of Hybrid Lambda. Hybrid lambda (X) E. coli DNA molecules were constructed using XgtXc DNA as the vehicle and fragments of DNA obtained by EcoRI endonuclease digestion of F factor carrying flagellar genes. The isolation of the F ...

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Section: Introductionmentioning

confidence: 99%

The identification of the mot gene product with Escherichia coli-lambda hybrids.

Silverman¹,

Matsumura²

1976

Proc. Natl. Acad. Sci. U.S.A.

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“…1). All bacterial flagella thus far studied, including those from C. crescentus (1), are composed of a filament with a cell-proximal flexible hook, which is attached to a transmembrane basal body (2)(3)(4). The C. crescentus flagellar filament is composed of two flagellins of 25,000 and 27,500 Mr (5,6).…”

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confidence: 99%

Methylation involved in chemotaxis is regulated during Caulobacter differentiation.

Shaw¹,

Gomes²,

Sweeney³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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Caulobacter crescentus carries a flagellum and is motile only during a limited time in its cell cycle. We have asked if the biochemical machinery that mediates chemotaxis exists coincident with the cell's structural ability to respond to a chemotactic signal. We first demonstrated that one function of the chemotaxis machinery, the ability to methylate the carboxyl side chains of a specific set of membrane proteins (methyl-accepting chemotaxis proteins, MCPs), is present in C. crescentus. This conclusion is based on the observations that (i) methionine auxotrophs starved of methionine can swim only in the forward direction (comparable to smooth swimming in the enteric bacteria), (ii) a specific set of membrane proteins was found to be methylated in vivo and the incorporated [3H]methyl groups were alkali sensitive, (iii) this same set of membrane proteins incorporated methyl groups from Sadenosylmethionine in vitro, and (iv) out of a total of eight generally nonchemotactic mutants, two were found to swim only in a forward direction and one of these lacked methyltransferase activity. Analysis of in vivo and in vitro methylation in synchronized cultures showed that the methylation reaction is lost when the flagellated swarmer cell differentiates into a stalked cell. In vivo methylation reappeared coincident with the biogenesis of the flagellum just prior to cell division. In vitro reconstitution experiments with heterologous cell fractions from different cell types showed that swarmer cells contain methyltransferase and their membranes can be methylated. However, newly differentiated stalked cells lack methyltransferase activity and membranes from these cells cannot accept methyl groups. These results demonstrate that MCP methylation is confined to that portion of the cell cycle when flagella are present.

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Biochemistry of Bacterial Flagella

Cited by 2 publications

References 23 publications

The identification of the mot gene product with Escherichia coli-lambda hybrids.

The identification of the mot gene product with Escherichia coli-lambda hybrids.

Methylation involved in chemotaxis is regulated during Caulobacter differentiation.

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