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Journal of Bacteriology, July 2002, p. 3871-3878, Vol. 184, No. 14
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.14.3871-3878.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

The PhlA Hemolysin from the Entomopathogenic Bacterium Photorhabdus luminescens Belongs to the Two-Partner Secretion Family of Hemolysins

Julien Brillard,1,{dagger} Eric Duchaud,2 Noël Boemare,1 Frank Kunst,2 and Alain Givaudan1*

Laboratoire EMIP, Université Montpellier II, IFR56, Institut National de la Recherche Agronomique (UMR 1133), 34095 Montpellier Cedex 5,1 Laboratoire de Génomique des Microorganismes Pathogènes, Institut Pasteur, 75724 Paris Cedex 15, France2

Received 18 December 2001/ Accepted 18 April 2002

Photorhabdus is an entomopathogenic bacterium symbiotically associated with nematodes of the family Heterorhabditidae. Bacterial hemolysins found in numerous pathogenic bacteria are often virulence factors. We describe here the nucleotide sequence and the molecular characterization of the Photorhabdus luminescens phlBA operon, a locus encoding a hemolysin which shows similarities to the Serratia type of hemolysins. It belongs to the two-partner secretion (TPS) family of proteins. In low-iron conditions, a transcriptional induction of the phlBA operon was observed by using the chloramphenicol acetyltransferase reporter gene, causing an increase in PhlA hemolytic activity compared to iron-rich media. A spontaneous phase variant of P. luminescens was deregulated in phlBA transcription. The phlA mutant constructed by allelic exchange remained highly pathogenic after injection in the lepidopteran Spodoptera littoralis, indicating that PhlA hemolysin is not a major virulence determinant. Using the gene encoding green fluorescent protein as a reporter, phlBA transcription was observed in hemolymph before insect death. We therefore discuss the possible role of PhlA hemolytic activity in the bacterium-nematode-insect interactions.

* Corresponding author. Mailing address: Laboratoire EMIP, CC101, INRA-Université Montpellier II (UMR 1133), Place E. Bataillon, 34095 Montpellier Cedex 5, France. Phone: 33-4-67144812. Fax: 33-4-67144679. E-mail: givaudan{at}crit.univ-montp2.fr.

{dagger} Present address: Unité de Biochimie Microbienne, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.

Journal of Bacteriology, July 2002, p. 3871-3878, Vol. 184, No. 14
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.14.3871-3878.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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