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Journal of Bacteriology, January 2004, p. 122-135, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.122-135.2004

Growth Phase-Dependent Regulation and Stringent Control of fis Are Conserved Processes in Enteric Bacteria and Involve a Single Promoter (fis P) in Escherichia coli

Prabhat Mallik, Timothy S. Pratt, Michael B. Beach,{dagger} Meranda D. Bradley, Jayanthi Undamatla,{ddagger} and Robert Osuna*

Department of Biological Sciences, University at Albany, SUNY, Albany, New York 12222

Received 24 July 2003/ Accepted 1 October 2003

The intracellular concentration of the Escherichia coli factor for inversion stimulation (Fis), a global regulator of transcription and a facilitator of certain site-specific DNA recombination events, varies substantially in response to changes in the nutritional environment and growth phase. Under conditions of nutritional upshift, fis is transiently expressed at very high levels, whereas under induced starvation conditions, fis is repressed by stringent control. We show that both of these regulatory processes operate on the chromosomal fis genes of the enterobacteria Klebsiella pneumoniae, Serratia marcescens, Erwinia carotovora, and Proteus vulgaris, strongly suggesting that the physiological role of Fis is closely tied to its transcriptional regulation in response to the nutritional environment. These transcriptional regulatory processes were previously shown to involve a single promoter (fis P) preceding the fis operon in E. coli. Recent work challenged this notion by presenting evidence from primer extension assays which appeared to indicate that there are multiple promoters upstream of fis P that contribute significantly to the expression and regulation of fis in E. coli. Thus, a rigorous analysis of the fis promoter region was conducted to assess the contribution of such additional promoters. However, our data from primer extension analysis, S1 nuclease mapping, ß-galactosidase assays, and in vitro transcription analysis all indicate that fis P is the sole E. coli fis promoter in vivo and in vitro. We further show how certain conditions used in the primer extension reactions can generate artifacts resulting from secondary annealing events that are the likely source of incorrect assignment of additional fis promoters.

* Corresponding author. Mailing address: Department of Biological Sciences, University at Albany, SUNY, 1400 Washington Avenue, Albany, NY 12222. Phone: (518) 437-4492. Fax: (518) 442-4767. E-mail: osuna{at}albany.edu.

{dagger} Present address: Department of Biology, Chemistry and Physics, Southern Polytechnic State University, Marietta, GA 30060.

{ddagger} Present address: Department of Molecular Medicine, Apollo Hospitals, Jubilee Hills, Hyderabad 500 033, India.

Journal of Bacteriology, January 2004, p. 122-135, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.122-135.2004




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