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Journal of Bacteriology, January 2005, p. 639-648, Vol. 187, No. 2
0021-9193/05/$08.00+0     doi:10.1128/JB.187.2.639-648.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Erwinia chrysanthemi Type III Secretion System Is Required for Multicellular Behavior

Mee-Ngan Yap,1 Ching-Hong Yang,2 Jeri D. Barak,3 Courtney E. Jahn,1 and Amy O. Charkowski1*

Department of Plant Pathology, University of Wisconsin—Madison, Madison,1 Department of Biological Sciences, University of Wisconsin—Milwaukee, Milwaukee, Wisconsin,2 Produce Safety and Microbiology Research Unit, U.S. Department of Agriculture-Agricultural Research Service, Albany, California3

Received 5 July 2004/ Accepted 27 August 2004

Enterobacterial animal pathogens exhibit aggregative multicellular behavior, which is manifested as pellicles on the culture surface and biofilms at the surface-liquid-air interface. Pellicle formation behavior requires production of extracellular polysaccharide, cellulose, and protein filaments, known as curli. Protein filaments analogous to curli are formed by many protein secretion systems, including the type III secretion system (TTSS). Here, we demonstrate that Erwinia chrysanthemi, which does not carry curli genes, requires the TTSS for pellicle formation. These data support a model where cellulose and generic protein filaments, which consist of either curli or TTSS-secreted proteins, are required for enterobacterial aggregative multicellular behavior. Using this assay, we found that hrpY, which encodes a two-component system response regulator homolog, is required for activity of hrpS, which encodes a {sigma}54-dependent enhancer-binding protein homolog. In turn, hrpS is required for activity of the sigma factor homolog hrpL, which activates genes encoding TTSS structural and secreted proteins. Pellicle formation was temperature dependent and pellicles did not form at 36°C, even though TTSS genes were expressed at this temperature. We found that cellulose is a component of the E. chrysanthemi pellicle but that pellicle formation still occurs in a strain with an insertion in a cellulose synthase subunit homolog. Since the TTSS, but not the cellulose synthase subunit, is required for E. chrysanthemi pellicle formation, this inexpensive assay can be used as a high throughput screen for TTSS mutants or inhibitors.

* Corresponding author. Mailing address: Department of Plant Pathology, 1630 Linden Dr., University of Wisconsin—Madison, Madison, WI 53706. Phone: (608) 262-7911. Fax: (608) 263-2626. E-mail: amyc{at}plantpath.wisc.edu.

Journal of Bacteriology, January 2005, p. 639-648, Vol. 187, No. 2
0021-9193/05/$08.00+0     doi:10.1128/JB.187.2.639-648.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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