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Laboratoire de Ghetique Moleculaire des Micro-organismes et des Interactions Cellulaires, CNRS-UMR 5577, INSA Bat 406i 2o Albert 69621 Villeurbanne, France
2Author for correspondence: Guy Condemine. Tel: + 33 472 43 80 88. Fax: + 33 472 43 87 14.e-mail: condemin@insa.insa-1yon.fr
ABSTRACT
SUMMARY: Secretion of pectate lyases and a cellulase occurs in winis chrysanthemi through a type II secretion machinery, the Out system. Proper insertion of the secretin OutD in the outer membrane requires the presence of Outs. Outs is an outer-membrane lipoprotein that interacts directly with OutD. Using ligand-blotting experiments, it has been shown that this interaction requires at least the 62 C-terminal amino acids of OutD. When this domain was added to the C-terminal extremity of the secreted pectate lyase PelD, the construct was stabilized by Outs but not inserted into the outer membrane. Thus, thisdomain is sufficient to interact with Outs but it is unable to confer the ability to be inserted into the outer membrane in the presence of Outs. A screen for outs mutants unable to secrete pectate lyases gave only mutants unable to properly localize OutD in the outer membrane and no mutant in the protection function. Thus, the interaction between Outs and OutD can probably not be abolished by the mutation of a single amino acid, and the insertion of OutD in the outer membrane may require additional proteins.
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