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Journal of Bacteriology, May 2001, p. 2918-2928, Vol. 183, No. 9
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.9.2918-2928.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Mapping Stress-Induced Changes in Autoinducer AI-2 Production in Chemostat-Cultivated Escherichia coli K-12

Matthew P. DeLisa,1,2 James J. Valdes,3 and William E. Bentley1,2,*

Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute,1 and Department of Chemical Engineering,2 University of Maryland, College Park, Maryland 20742, and U.S. Army Edgewood Chemical Biological Center, Aberdeen Proving Ground, Maryland 210103

Received 8 November 2000/Accepted 6 February 2001

Numerous gram-negative bacteria employ a cell-to-cell signaling mechanism, termed quorum sensing, for controlling gene expression in response to population density. Recently, this phenomenon has been discovered in Escherichia coli, and while pathogenic E. coli utilize quorum sensing to regulate pathogenesis (i.e., expression of virulence genes), the role of quorum sensing in nonpathogenic E. coli is less clear, and in particular, there is no information regarding the role of quorum sensing during the overexpression of recombinant proteins. The production of autoinducer AI-2, a signaling molecule employed by E. coli for intercellular communication, was studied in E. coli W3110 chemostat cultures using a Vibrio harveyi AI-2 reporter assay (M. G. Surrette and B. L. Bassler, Proc. Natl. Acad. Sci. USA 95:7046-7050, 1998). Chemostat cultures enabled a study of AI-2 regulation through steady-state and transient responses to a variety of environmental stimuli. Results demonstrated that AI-2 levels increased with the steady-state culture growth rate. In addition, AI-2 increased following pulsed addition of glucose, Fe(III), NaCl, and dithiothreitol and decreased following aerobiosis, amino acid starvation, and isopropyl-beta -D-thiogalactopyranoside-induced expression of human interleukin-2 (hIL-2). In general, the AI-2 responses to several perturbations were indicative of a shift in metabolic activity or state of the cells induced by the individual stress. Because of our interest in the expression of heterologous proteins in E. coli, the transcription of four quorum-regulated genes and 20 stress genes was mapped during the transient response to induced expression of hIL-2. Significant regulatory overlap was revealed among several stress and starvation genes and known quorum-sensing genes.

* Corresponding author. Mailing address: Center for Agricultural Biotechnology, University of Maryland Biotechnology Institute, University of Maryland, College Park, MD 20742. Phone: (301) 405-4321. Fax: (301) 314-9075. E-mail: bentley{at}eng.umd.edu.

Journal of Bacteriology, May 2001, p. 2918-2928, Vol. 183, No. 9
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.9.2918-2928.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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