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Originally published In Press as doi:10.1074/jbc.M101129200 on February 9, 2001

J. Biol. Chem., Vol. 276, Issue 20, 17190-17198, May 18, 2001
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Identification of an ATP-binding Cassette Transporter for Export of the O-antigen across the Inner Membrane in Rhizobium etli Based on the Genetic, Functional, and Structural Analysis of an lps Mutant Deficient in O-antigen*

Inge LerougeDagger , Toon LaeremansDagger , Christel VerrethDagger , Jos VanderleydenDagger §, Caroline Van SoomDagger §||, Andrea Tobin**, and Russell W. Carlson**Dagger Dagger

From the Dagger  Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, Heverlee B-3001, Belgium and the ** Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602

For O-antigen lipopolysaccharide (LPS) synthesis in bacteria, transmembrane migration of undecaprenyl pyrophosphate-bound O-antigen oligosaccharide subunits or polysaccharide occurs before ligation to the core region of the LPS molecule. In this study, we identified by mutagenesis an ATP-binding cassette transporter in Rhizobium etli CE3 that is likely responsible for the translocation of the O-antigen across the inner plasma membrane. Mutant FAJ1200 LPS lacks largely the O-antigen, as shown by SDS-polyacrylamide gel electrophoresis and confirmed by immunoblot analysis. Furthermore, LPS isolated from FAJ1200 is totally devoid of any O-chain glycosyl residues and contains only those glycosyl residues that can be expected for the inner core region. The membrane component and the cytoplasmic ATP-binding component of the ATP-binding cassette transporter are encoded by wzm and wzt, respectively. The Tn5 transposon in mutant FAJ1200 is inserted in the wzm gene. This mutation resulted in an Inf- phenotype in bean plants.


* This work was supported in part by National Institutes of Health Grant GM39583 (to R. W. C.) and Department of Energy Grant DE-FG09-93ER20097 (to the Complex Carbohydrate Research Center).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF182824.

§ Supported by "Fonds Wetenschappelijk Onderzoek-Vlaanderen" and the Flemish Government (Geconcerteerde Onderzoeks Actie/98/Vanderleyden).

|| Postdoctoral Fellow of the Belgian Fund for Scientific Research.

Dagger Dagger To whom correspondence should be addressed. Tel.: 706-542-4439; Fax: 706-542-4412; E-mail: RCARLSON@ccrc.uga.edu.

To whom correspondence should be addressed. Tel.: 32-16-32-1631; Fax: 32-16-32-1966; E-mail: Jos.Vanderleyden@agr.kuleuven.ac.be.


Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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