HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Rawlinson, E. L. A. Articles by Skaugen, M. Search for Related Content PubMed PubMed Citation Articles by Rawlinson, E. L. A. Articles by Skaugen, M. Agricola Articles by Rawlinson, E. L. A. Articles by Skaugen, M.

Identification of the DNA-binding site of the Rgg-like regulator LasX within the lactocin S promoter region

Elizabeth L. Andersen Rawlinson

Morten Skaugen

Laboratory of Microbial Gene Technology, PO Box 5003, Norwegian University of Life Sciences, N-1432 Ås, Norway

Correspondence
Morten Skaugen
morten.skaugen{at}umb.no

LasX regulates the transcription of the divergent operons lasXY and lasA–W, which specify the production of lactocin S in Lactobacillus sakei L45. Using histidine-tagged LasX, and a DNA fragment containing the complete intergenic lasA–lasX region, electrophoresis mobility-shift (EMSA) analyses were employed to demonstrate that LasX binds to the lasA–lasX intergenic DNA. Two direct heptanucleotide motifs directly upstream of P_lasA–W, and a third imperfect copy of this motif, overlapping the –10 element of P_lasA–W, were identified as possible LasX-binding sites. To assess the role of the direct repeats in the binding of LasX to the intergenic lasA–lasX region, binding experiments were performed using DNA probes with different combinations of the repeats, and with arbitrarily chosen repeat substitutions. The result of these experiments demonstrated that only the middle repeat was required for the binding of LasX to the las-promoter region. This observation correlated with the results of subsequent reporter-gene analyses, thereby weakening the hypothesis of the involvement of the direct repeats in LasX-mediated transcription regulation. By analysing the ability of LasX to bind successively shortened derivatives of the original intergenic fragment, a tentative 19 bp minimum LasX-binding site was identified.

Present address: Norwegian Defence Research Establishment, PO Box 25, N-2027 Kjeller, Norway.

Present address: Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, PO Box 5003, N-1432 Ås-NLH, Norway.

This article has been cited by other articles:

				I. F. Nes, D. B. Diep, and H. Holo Bacteriocin Diversity in Streptococcus and Enterococcus J. Bacteriol., February 15, 2007; 189(4): 1189 - 1198. [Full Text] [PDF]

				J. A. Loughman and M. G. Caparon Contribution of Invariant Residues to the Function of Rgg Family Transcription Regulators J. Bacteriol., January 15, 2007; 189(2): 650 - 655. [Abstract] [Full Text] [PDF]

				A. V. Dmitriev, E. J. McDowell, K. V. Kappeler, M. A. Chaussee, L. D. Rieck, and M. S. Chaussee The Rgg Regulator of Streptococcus pyogenes Influences Utilization of Nonglucose Carbohydrates, Prophage Induction, and Expression of the NAD-Glycohydrolase Virulence Operon. J. Bacteriol., October 1, 2006; 188(20): 7230 - 7241. [Abstract] [Full Text] [PDF]

				M. A. Chaussee, E. J. McDowell, L. D. Rieck, E. A. Callegari, and M. S. Chaussee Proteomic analysis of a penicillin-tolerant rgg mutant strain of Streptococcus pyogenes J. Antimicrob. Chemother., October 1, 2006; 58(4): 752 - 759. [Abstract] [Full Text] [PDF]