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Infection and Immunity, March 2009, p. 1208-1215, Vol. 77, No. 3
0019-9567/09/$08.00+0     doi:10.1128/IAI.01006-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Regulation of the Vibrio vulnificus hupA Gene by Temperature Alteration and Cyclic AMP Receptor Protein and Evaluation of Its Role in Virulence{triangledown}

Man Hwan Oh,{dagger} Sung Min Lee,{dagger},{ddagger} Dong Hwan Lee, and Sang Ho Choi*

National Research Laboratory of Molecular Microbiology and Toxicology, Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, South Korea

Received 6 August 2008/ Returned for modification 23 October 2008/ Accepted 29 December 2008

Availability of free iron is extremely limited in the mammalian host, and the acquisition of iron in the host is essential for successful infection by pathogenic bacteria. Expression of many genes involved in acquiring iron is regulated in response to the level of iron availability, and iron regulation is mediated by Fur. In this study, cellular levels of Vibrio vulnificus HupA, a heme receptor protein, and the hupA transcript were found to increase in cells grown at 40°C compared to cells grown at 30°C. The results suggested that change in growth temperature, in addition to iron availability, is an environmental cue controlling the expression of the hupA gene. The influence of global regulatory proteins on the expression of hupA was examined, and the cyclic AMP receptor protein (CRP) was found to activate the expression of hupA at the transcriptional level. CRP exerts its effects by directly binding to DNA upstream of the hupA promoter PhupA, and a CRP binding site, centered at 174 bp upstream of the transcription start site, was identified by a DNase I protection assay. Finally, a hupA mutant showed reduced virulence in mice and in tissue cultures, in which growth of the hupA mutant was impaired, indicating that HupA of V. vulnificus is essential for survival and multiplication during infection.

* Corresponding author. Mailing address: Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, South Korea. Phone: 82-2-880-4857. Fax: 82-2-873-5095. E-mail: choish{at}snu.ac.kr

{triangledown} Published ahead of print on 12 January 2009.

Editor: V. J. DiRita

{dagger} M.H.O. and S.M.L. contributed equally to this work.

{ddagger} Present address: Food Sanitation Council, Codex Office, Korea Food and Drug Administration, Seoul 122-704, South Korea.

Infection and Immunity, March 2009, p. 1208-1215, Vol. 77, No. 3
0019-9567/09/$08.00+0     doi:10.1128/IAI.01006-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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