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Journal of Bacteriology, February 2000, p. 821-824, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification by Genetic Suppression of Escherichia coli TolB Residues Important for TolB-Pal Interaction

Marie-Céline Ray, Pierre Germon, Anne Vianney, Raymond Portalier, and Jean Claude Lazzaroni*

Unité de Microbiologie et Génétique, UMR 5577, CNRS-INSA-Université Lyon I, F-69622 Villeurbanne, France

Received 12 July 1999/Accepted 8 November 1999

The Tol-Pal system of Escherichia coli is involved in maintaining outer membrane stability. Mutations in tolQ, tolR, tolA, tolB, or pal genes result in sensitivity to bile salts and the leakage of periplasmic proteins. Moreover, some of the tol genes are necessary for the entry of group A colicins and the DNA of filamentous bacteriophages. TolQ, TolR, and TolA are located in the cytoplasmic membrane where they interact with each other via their transmembrane domains. TolB and Pal form a periplasmic complex near the outer membrane. We used suppressor genetics to identify the regions important for the interaction between TolB and Pal. Intragenic suppressor mutations were characterized in a domain of Pal that was shown to be involved in interactions with TolB and peptidoglycan. Extragenic suppressor mutations were located in tolB gene. The C-terminal region of TolB predicted to adopt a beta -propeller structure was shown to be responsible for the interaction of the protein with Pal. Unexpectedly, none of the suppressor mutations was able to restore a correct association between Pal and peptidoglycan, suggesting that interactions between Pal and other components such as TolB may also be important for outer membrane stability.

* Corresponding author. Mailing address: Unité de Microbiologie et Génétique, UMR 5577, bât 405, F-69622 Villeurbanne Cedex, France. Phone: (33) 472 43 13 67. Fax: (33) 472 43 26 86. E-mail: lazzaroni{at}biomserv.univ-lyon1.fr.

Journal of Bacteriology, February 2000, p. 821-824, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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