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J. Biol. Chem., Vol. 281, Issue 46, 34775-34784, November 17, 2006

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Transcriptional Regulatory Cascade for Elastase Production in Vibrio vulnificus

LuxO ACTIVATES luxT EXPRESSION AND LuxT REPRESSES smcR EXPRESSION^*

Jong-Bok Roh¹, Mi-Ae Lee¹, Hyun-Jung Lee, Sung-Min Kim, Yona Cho, You-Jin Kim, Yeong-Jae Seok, Soon-Jung Park^¶, and Kyu-Ho Lee²

From the Department of Environmental Science, Hankuk University of Foreign Studies, Yongin, Kyunggi-Do 449-791, the Department of Biological Sciences, Seoul National University, Seoul 151 742, and the ^¶Department of Parasitology, The Brain Korea 21 Project, Yonsei University, College of Medicine, Seoul, 133-791, South Korea

Vibrio vulnificus causes diseases through actions of various virulence factors, including the elastase encoded by the vvpE gene. Through transposon mutagenesis of V. vulnificus, vvpE expression was shown to be increased by luxO mutation. Since the vvpE gene is known to be positively regulated by SmcR via direct binding to the vvpE promoter, the role of LuxO in smcR expression was investigated. The luxAB-transcriptional fusions containing different lengths of the smcR promoter region indicated that the smcR transcription was negatively regulated by LuxO and that a specific upstream region of the smcR gene was required for this repression. Since LuxO is a known member of positive regulators, the negative regulation of smcR transcription by LuxO prompted us to identify the factor(s) linking LuxO and smcR transcription. LuxT was isolated in a ligand fishing experiment using the smcR upstream region as bait, and smcR expression was increased by luxT mutation. Recombinant LuxT bound to a specific upstream region of the smcR gene, –154 to –129 relative to the smcR transcription start site. The expression of luxT was positively regulated by LuxO, and the luxT promoter region contained a putative LuxO-binding site. Mutagenesis of the LuxO-binding site in the luxT promoter region resulted in a loss of transcriptional control by LuxO. Therefore, this study demonstrates a transcriptional regulatory cascade for elastase production, where LuxO activates luxT transcription and LuxT represses smcR transcription.

Received for publication, August 16, 2006 , and in revised form, September 5, 2006.

^* This study was supported by Korea Science and Engineering Foundation, Republic of Korea, Grant R01-2004-000-10046-0 (to S. H. C. and K.-H. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) DQ778302 [GenBank] .

The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. 1.

¹ These two authors contributed equally to this work.

² To whom correspondence should be addressed: Dept. of Environmental Science, Hankuk University of Foreign Studies, Wangsan-Li, Mohyun-Myun, Yongin, Kyunggi-Do 449-791, Korea. Tel.: 82-31-330-4039; Fax: 82-31-330-4529; E-mail: khlee{at}hufs.ac.kr.

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