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J. Biol. Chem., Vol. 276, Issue 20, 17190-17198, May 18, 2001
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From the 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 The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF182824.
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*
,
,
,
§¶,
§
,
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
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.
Postdoctoral Fellow of the Belgian Fund for Scientific Research.

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.
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