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Journal of Bacteriology, March 2003, p. 1951-1957, Vol. 185, No. 6
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.6.1951-1957.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of Catabolite Repression as a Physiological Regulator of Biofilm Formation by Bacillus subtilis by Use of DNA Microarrays

Nicola R. Stanley,1 Robert A. Britton,2,{dagger} Alan D. Grossman,2 and Beth A. Lazazzera1*

Department of Microbiology, Immunology and Molecular Genetics, University of California—Los Angeles, Los Angeles, California 90095,1 Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 021392

Received 17 October 2002/ Accepted 20 December 2002

Biofilms are structured communities of cells that are encased in a self-produced polymeric matrix and are adherent to a surface. Many biofilms have a significant impact in medical and industrial settings. The model gram-positive bacterium Bacillus subtilis has recently been shown to form biofilms. To gain insight into the genes involved in biofilm formation by this bacterium, we used DNA microarrays representing >99% of the annotated B. subtilis open reading frames to follow the temporal changes in gene expression that occurred as cells transitioned from a planktonic to a biofilm state. We identified 519 genes that were differentially expressed at one or more time points as cells transitioned to a biofilm. Approximately 6% of the genes of B. subtilis were differentially expressed at a time when 98% of the cells in the population were in a biofilm. These genes were involved in motility, phage-related functions, and metabolism. By comparing the genes differentially expressed during biofilm formation with those identified in other genomewide transcriptional-profiling studies, we were able to identify several transcription factors whose activities appeared to be altered during the transition from a planktonic state to a biofilm. Two of these transcription factors were Spo0A and sigma-H, which had previously been shown to affect biofilm formation by B. subtilis. A third signal that appeared to be affecting gene expression during biofilm formation was glucose depletion. Through quantitative biofilm assays and confocal scanning laser microscopy, we observed that glucose inhibited biofilm formation through the catabolite control protein CcpA.

* Corresponding author. Mailing address: Department of Microbiology, Immunology and Molecular Genetics, University of California—Los Angeles, 1602 Molecular Sciences Building, 405 Hilgard Ave., Los Angeles, CA 90095. Phone: (310) 794-4804. Fax: (310) 206-5231. E-mail: bethl{at}microbio.ucla.edu.

{dagger} Present address: Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48823.

Journal of Bacteriology, March 2003, p. 1951-1957, Vol. 185, No. 6
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.6.1951-1957.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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