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Journal of Bacteriology, February 2004, p. 611-622, Vol. 186, No. 3
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.3.611-622.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

In Vivo Expression of the Mannose-Resistant Fimbriae of Photorhabdus temperata K122 during Insect Infection

L. M. Meslet-Cladiere,1 A. Pimenta,1,2 E. Duchaud,3 I. B. Holland,1 and M. A. Blight1*

Institut de Génétique et Microbiologie, CNRS UMR 8621, Laboratoire de Pathogenèse Comparée, Université Paris XI, 91405 Orsay Cedex,1 Laboratoire ERRMECe, Group d'Interactions Cellulaires, Université de Cergy-Pontoise, 95302 Cergy-Pontoise Cedex,2 Atelier de BioInformatique (ABI), 75252 Paris Cedex 05, France3

Received 12 August 2003/ Accepted 24 October 2003

Photorhabdus temperata K122 is an entomopathogenic bacterium symbiotically associated with nematodes of the family Heterorhabditidae. Surface fimbriae are important for the colonization of many pathogenic bacteria, and here we report the nucleotide sequence and analysis of the expression of a 12-kbp fragment encoding the mannose-resistant fimbriae of P. temperata (mrf). The mrf gene cluster contains 11 genes with an organization similar to that of the mrp locus from Proteus mirabilis. mrfI (encoding a putative recombinase) and mrfA (encoding pilin), the first gene in an apparent operon of nine other genes, are expressed from divergent promoters. The mrfI-mrfA intergenic region contains inverted repeats flanking the mrfA promoter. This region was shown to be capable of inversion, consistent with an ON/OFF regulation of the operon. In in vitro liquid cultures, both orientations were detected. Nevertheless, when we analyzed the expression of all of the genes in the mrf locus by semiquantitative reverse transcription-PCR during infection of Galleria mellonella (greater wax moth) larvae, expression of mrfA was not detected until 25 h postinfection, preceding the death of the larvae at 32 h. In contrast, mrfJ (a putative inhibitor of flagellar synthesis) was expressed throughout infection. Expression of mrfI was also detected only late in infection (25 to 30 h), indicating a possible increase in inversion frequency at this stage. In both in vitro liquid cultures and in vivo larval infections, the distal genes of the operon were expressed at substantially lower levels than mrfA. These results indicate the complex regulation of the mrf cluster during infection.

* Corresponding author. Mailing address: Institut de Génétique et Microbiologie, CNRS UMR 8621, Laboratoire de Pathogenèse Comparée, BÂtiment 360 et 409, Université Paris XI, 91405 Orsay Cedex, France. Phone: (33 1) 69158168. Fax: (33 1) 69156334. E-mail: mark.blight{at}igmors.u-psud.fr.

Journal of Bacteriology, February 2004, p. 611-622, Vol. 186, No. 3
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.3.611-622.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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