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Infection and Immunity, April 2009, p. 1543-1552, Vol. 77, No. 4
0019-9567/09/$08.00+0     doi:10.1128/IAI.00949-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

The CsgA and Lpp Proteins of an Escherichia coli O157:H7 Strain Affect HEp-2 Cell Invasion, Motility, and Biofilm Formation {triangledown}

Gaylen A. Uhlich,1* Nereus W. Gunther IV,1 Darrell O. Bayles,2 and Derek A. Mosier3

Microbial Food Safety Research Unit, Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, Pennsylvania,1 Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Ames, Iowa,2 Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, 1600 Denison Ave., Manhattan, Kansas3

Received 29 July 2008/ Returned for modification 13 September 2008/ Accepted 17 January 2009

In Escherichia coli O157:H7 strain ATCC 43895, a guanine-to-thymine transversion in the csgD promoter created strain 43895OR. Strain 43895OR produces an abundant extracellular matrix rich in curli fibers, forms biofilms on solid surfaces, invades cultured epithelial cells, and is more virulent in mice than strain 43895. In this study we compared the formic acid-soluble proteins expressed by strains 43895OR and 43895 using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and identified two differentially expressed proteins. A 17-kDa protein unique to strain 43895OR was identified from matrix-assisted laser desorption ionization-time of flight analysis combined with mass spectrometry (MS) and tandem MS (MS/MS) as the curli subunit encoded by csgA. A <10-kDa protein, more highly expressed in strain 43895, was identified as the Lpp lipoprotein. Mutants of strain 43895OR with disruption of lpp, csgA, or both lpp and csgA were created and tested for changes in phenotype and function. The results of this study show that both Lpp and CsgA contribute to the observed colony morphology, Congo red binding, motility, and biofilm formation. We also show that both CsgA and Lpp are required by strain 43895OR for the invasion of cultured HEp-2 cells. These studies suggest that in strain 43895OR, the murein lipoprotein Lpp indirectly regulates CsgA expression through the CpxAR system by a posttranscriptional mechanism.

* Corresponding author. Mailing address: Eastern Regional Research Center, ARS, USDA, 600 E. Mermaid Lane, Wyndmoor, PA 19038. Phone: (215) 233-6740. Fax: (215) 233-6581. E-mail: gaylen.uhlich{at}ars.usda.gov

{triangledown} Published ahead of print on 29 January 2009.

Editor: A. Camilli

Infection and Immunity, April 2009, p. 1543-1552, Vol. 77, No. 4
0019-9567/09/$08.00+0     doi:10.1128/IAI.00949-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Uhlich, G. A. (2009). KatP contributes to OxyR-regulated hydrogen peroxide resistance in Escherichia coli serotype O157 : H7. Microbiology 155: 3589-3598 [Abstract] [Full Text]  


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