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Infection and Immunity, November 2008, p. 5016-5027, Vol. 76, No. 11
0019-9567/08/$08.00+0     doi:10.1128/IAI.00314-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Functional Mapping of YadA- and Ail-Mediated Binding of Human Factor H to Yersinia enterocolitica Serotype O:3{triangledown}

Marta Biedzka-Sarek,1 Saara Salmenlinna,1 Markus Gruber,2 Andrei N. Lupas,2 Seppo Meri,1,3 and Mikael Skurnik1,3*

Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, 00014 Helsinki, Finland,1 Department of Protein Evolution, Max Planck Institute for Developmental Biology, D-72076 Tübingen, Germany,2 Helsinki University Central Hospital Laboratory Diagnostics, Helsinki, Finland3

Received 10 March 2008/ Returned for modification 24 April 2008/ Accepted 16 August 2008

Yersinia enterocolitica is an enteric pathogen that exploits diverse means to survive in the human host. Upon Y. enterocolitica entry into the human host, bacteria sense and respond to variety of signals, one of which is the temperature. Temperature in particular has a profound impact on Y. enterocolitica gene expression, as most of its virulence factors are expressed exclusively at 37°C. These include two outer membrane proteins, YadA and Ail, that function as adhesins and complement resistance (CR) factors. Both YadA and Ail bind the functionally active complement alternative pathway regulator factor H (FH). In this study, we characterized regions on both proteins involved in CR and the interaction with FH. Twenty-eight mutants having short (7 to 41 amino acids) internal deletions within the neck and stalk of YadA and two complement-sensitive site-directed Ail mutants were constructed to map the CR and FH binding regions of YadA and Ail. Functional analysis of the YadA mutants revealed that the stalk of YadA is required for both CR and FH binding and that FH appears to target several conformational and discontinuous sites of the YadA stalk. On the other hand, the complement-sensitive Ail mutants were not affected in FH binding. Our results also suggested that Ail- and YadA-mediated CR does not depend solely on FH binding.

* Corresponding author. Mailing address: Department of Bacteriology and Immunology, Haartman Institute, P.O. Box 21, 00014 University of Helsinki, Helsinki, Finland. Phone: 358-9-19126264. Fax: 358-9-19126382. E-mail: mikael.skurnik{at}helsinki.fi

{triangledown} Published ahead of print on 2 September 2008.

Editor: J. B. Bliska

Infection and Immunity, November 2008, p. 5016-5027, Vol. 76, No. 11
0019-9567/08/$08.00+0     doi:10.1128/IAI.00314-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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