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Identification of Genes Required for Synthesis of the Adhesive Holdfast in Caulobacter crescentus

Chris S. Smith, Aaron Hinz, Diane Bodenmiller, David E. Larson, Yves V. Brun
Chris S. Smith
Department of Biology, Indiana University, Bloomington, Indiana 47405
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Aaron Hinz
Department of Biology, Indiana University, Bloomington, Indiana 47405
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Diane Bodenmiller
Department of Biology, Indiana University, Bloomington, Indiana 47405
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David E. Larson
Department of Biology, Indiana University, Bloomington, Indiana 47405
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Yves V. Brun
Department of Biology, Indiana University, Bloomington, Indiana 47405
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  • For correspondence: ybrun@bio.indiana.edu
DOI: 10.1128/JB.185.4.1432-1442.2003
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ABSTRACT

Adhesion to both abiotic and biotic surfaces by the gram-negative prothescate bacterium Caulobacter crescentus is mediated by a polar organelle called the “holdfast,” which enables the bacterium to form stable monolayer biofilms. The holdfast, a complex polysaccharide composed in part of N-acetylglucosamine, localizes to the tip of the stalk (a thin cylindrical extension of the cell wall and membranes). We report here the isolation of adhesion mutants with transposon insertions in an uncharacterized gene cluster involved in holdfast biogenesis (hfs) as well as in previously identified polar development genes (podJ and pleC), and the holdfast attachment genes (hfa). Clean deletions of three of the four genes in the hfs gene cluster (hfsDAB) resulted in a severe holdfast biogenesis phenotype. These mutants do not bind to surfaces or to a fluorescently labeled lectin, specific for N-acetylglucosamine. Transmission electron microscopy indicated that the hfsDAB mutants fail to synthesize a holdfast at the stalk tip. The predicted hfs gene products have significant sequence similarity to proteins necessary for exopolysaccharide export in gram-negative bacteria. HfsA has sequence similarity to GumC from Xanthomonas campestris, which is involved in exopolysaccharide export in the periplasm. HfsD has sequence similarity to Wza from Escherichia coli, an outer membrane protein involved in secretion of polysaccharide through the outer membrane. HfsB is a novel protein involved in holdfast biogenesis. These data suggest that the hfs genes play an important role in holdfast export.

  • Copyright © 2003 American Society for Microbiology
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Identification of Genes Required for Synthesis of the Adhesive Holdfast in Caulobacter crescentus
Chris S. Smith, Aaron Hinz, Diane Bodenmiller, David E. Larson, Yves V. Brun
Journal of Bacteriology Feb 2003, 185 (4) 1432-1442; DOI: 10.1128/JB.185.4.1432-1442.2003

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Identification of Genes Required for Synthesis of the Adhesive Holdfast in Caulobacter crescentus
Chris S. Smith, Aaron Hinz, Diane Bodenmiller, David E. Larson, Yves V. Brun
Journal of Bacteriology Feb 2003, 185 (4) 1432-1442; DOI: 10.1128/JB.185.4.1432-1442.2003
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KEYWORDS

bacterial adhesion
Bacterial Proteins
Caulobacter crescentus
mutation
Organelles

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