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Journal of Bacteriology, January 1999, p. 382-388, Vol. 181, No. 2
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Assembly of XcpR in the Cytoplasmic Membrane Is Required for Extracellular Protein Secretion in Pseudomonas aeruginosa

Geneviève Ball, Virginie Chapon-Hervé, Sophie Bleves,dagger Gérard Michel, and Marc Bally*

Laboratoire d'Ingénierie des Systèmes Macromoléculaires, Centre National de la Recherche Scientifique, 13402 Marseille Cedex 20, France

Received 19 June 1998/Accepted 26 October 1998

A broad range of extracellular proteins secreted by Pseudomonas aeruginosa use the type II or general secretory pathway (GSP) to reach the medium. This pathway requires the expression of at least 12 xcp gene products. XcpR, a putative nucleotide-binding protein, is essential for the secretion process across the outer membrane even though the protein contains no hydrophobic sequence that could target or anchor it to the bacterial envelope. For a better understanding of the relationship between XcpR and the other Xcp proteins which are located in the envelope, we have studied its subcellular localization. In a wild-type P. aeruginosa strain, XcpR was found associated with the cytoplasmic membrane. This association depends on the presence of the XcpY protein, which also appears to be necessary for XcpR stability. Functional complementation of an xcpY mutant required the XcpY protein to be expressed at a low level. Higher expression precluded the complementing activity of XcpY, although membrane association of XcpR was restored. This behavior suggested that an excess of free XcpY might interfere with the secretion by formation of inactive XcpR-XcpY complexes which cannot properly interact with their natural partners in the secretion machinery. These data show that a precise stoichiometric ratio between several components may be crucial for the functioning of the GSP.


* Corresponding author. Mailing address: Laboratoire d'Ingénierie des Systèmes Macromoléculaires, Centre National de la Recherche Scientifique, 31 Chemin Joseph-Aiguier, 13402 Marseille Cedex 20, France. Phone: 33 (0)491-164487. Fax: 33 (0)491-712124. E-mail: bally{at}ibsm.cnrs-mrs.fr.

dagger Present address: International Institute of Cellular and Molecular Pathology and Faculté de Medecine, Université Catholique de Louvain, UCL 74-49, B-1200 Brussels, Belgium.


Journal of Bacteriology, January 1999, p. 382-388, Vol. 181, No. 2
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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