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Journal of Bacteriology, February 2008, p. 1097-1107, Vol. 190, No. 3
0021-9193/08/$08.00+0     doi:10.1128/JB.00516-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Campylobacter jejuni Biofilms Up-Regulated in the Absence of the Stringent Response Utilize a Calcofluor White-Reactive Polysaccharide{triangledown}

Meghan K. McLennan,1,{dagger} Danielle D. Ringoir,1,{dagger} Emilisa Frirdich,1,{ddagger} Sarah L. Svensson,1,{ddagger} Derek H. Wells,2 Harold Jarrell,3 Christine M. Szymanski,3 and Erin C. Gaynor1*

Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada,1 Division of Infectious Diseases, University of California San Francisco, San Francisco, California,2 Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada3

Received 4 April 2007/ Accepted 30 October 2007

The enteric pathogen Campylobacter jejuni is a highly prevalent yet fastidious bacterium. Biofilms and surface polysaccharides participate in stress survival, transmission, and virulence in C. jejuni; thus, the identification and characterization of novel genes involved in each process have important implications for pathogenesis. We found that C. jejuni reacts with calcofluor white (CFW), indicating the presence of surface polysaccharides harboring β1-3 and/or β1-4 linkages. CFW reactivity increased with extended growth, under 42°C anaerobic conditions, and in a {Delta}spoT mutant defective for the stringent response (SR). Conversely, two newly isolated dim mutants exhibited diminished CFW reactivity as well as growth and serum sensitivity differences from the wild type. Genetic, biochemical, and nuclear magnetic resonance analyses suggested that differences in CFW reactivity between wild-type and {Delta}spoT and dim mutant strains were independent of well-characterized lipooligosaccharides, capsular polysaccharides, and N-linked polysaccharides. Targeted deletion of carB downstream of the dim13 mutation also resulted in CFW hyporeactivity, implicating a possible role for carbamoylphosphate synthase in the biosynthesis of this polysaccharide. Correlations between biofilm formation and production of the CFW-reactive polymer were demonstrated by crystal violet staining, scanning electron microscopy, and confocal microscopy, with the C. jejuni {Delta}spoT mutant being the first SR mutant in any bacterial species identified as up-regulating biofilms. Together, these results provide new insight into genes and processes important for biofilm formation and polysaccharide production in C. jejuni.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of British Columbia, 2350 Health Sciences Mall, Vancouver, British Columbia, Canada V6T 1Z3. Phone: (604) 822-2710. Fax: (604) 822-6041. E-mail: egaynor{at}interchange.ubc.ca

{triangledown} Published ahead of print on 9 November 2007.

{dagger} M.K.M. and D.D.R. contributed equally to this work.

{ddagger} E.F. and S.L.S. contributed equally to this work.

Journal of Bacteriology, February 2008, p. 1097-1107, Vol. 190, No. 3
0021-9193/08/$08.00+0     doi:10.1128/JB.00516-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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