HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 51, 51347-51359, December 19, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/51/51347    most recent M309016200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Forsberg, L. S. Articles by Carlson, R. W. Search for Related Content PubMed PubMed Citation Articles by Forsberg, L. S. Articles by Carlson, R. W. Social Bookmarking                      What's this?

Genetic Locus and Structural Characterization of the Biochemical Defect in the O-Antigenic Polysaccharide of the Symbiotically Deficient Rhizobium etli Mutant, CE166

REPLACEMENT OF N-ACETYLQUINOVOSAMINE WITH ITS HEXOSYL-4-ULOSE PRECURSOR^*

L. Scott Forsberg, K. Dale Noel, Jodie Box, and Russell W. Carlson¶

From the Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602 and the Department of Biological Sciences, Marquette University, Milwaukee, Wisconsin 53233

The O-antigen polysaccharide (OPS) of Rhizobium etli CE3 lipopolysaccharide (LPS) is linked to the core oligosaccharide via an N-acetylquinovosaminosyl (QuiNAc) residue. A mutant of CE3, CE166, produces LPS with reduced amounts of OPS, and a suppressed mutant, CE166, produces LPS with nearly normal OPS levels. Both mutants are deficient in QuiNAc production. Characterization of OPS from CE166 and CE166 showed that QuiNAc was replaced by its 4-keto derivative, 2-acetamido-2,6-dideoxyhexosyl-4-ulose. The identity of this residue was determined by NMR and mass spectrometry, and by gas chromatography-mass spectrometry analysis of its 2-acetamido-4-deutero-2,6-dideoxyhexosyl derivatives produced by reduction of the 4-keto group using borodeuteride. Mass spectrometric and methylation analyses showed that the 2-acetamido-2,6-dideoxyhexosyl-4-ulosyl residue was 3-linked and attached to the core-region external Kdo III residue of the LPS, the same position as that of QuiNAc in the CE3 LPS. DNA sequencing revealed that the transposon insertion in strain CE166 was located in an open reading frame whose predicted translation product, LpsQ, falls within a large family of predicted open reading frames, which includes biochemically characterized members that are sugar epimerases and/or reductases. A hypothesis to be tested in future work is that lpsQ encodes UDP-2-acetamido-2,6-dideoxyhexosyl-4-ulose reductase, the second step in the synthesis of UDP-QuiNAc from UDP-GlcNAc.

Received for publication, August 14, 2003 , and in revised form, October 1, 2003.

^* This work was supported in part by National Institutes of Health Grant GM39583 (to R. W. C.), by Department of Energy (DOE) Grant DE-FG09-93ER20097 (to the Complex Carbohydrate Research Center), and by DOE Grant DE-FG02-98ER20307 (to K. D. N.). The costs of publication of this article were defrayed in part by the payment of page charges. This 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 GenBank^TM/EBI Data Bank with accession number(s) AY391267.

^¶ To whom correspondence should be addressed. Tel.: 706-542-4439; Fax: 706-542-4412; E-mail: rcarlson{at}ccrc.uga.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Lukasiewicz, T. Niedziela, W. Jachymek, L. Kenne, and C. Lugowski Two Kdo-Heptose Regions Identified in Hafnia alvei 32 Lipopolysaccharide: the Complete Core Structure and Serological Screening of Different Hafnia O Serotypes J. Bacteriol., January 15, 2009; 191(2): 533 - 544. [Abstract] [Full Text] [PDF]

				L. S. Forsberg and R. W. Carlson Structural Characterization of the Primary O-antigenic Polysaccharide of the Rhizobium leguminosarum 3841 Lipopolysaccharide and Identification of a New 3-Acetimidoylamino-3-deoxyhexuronic Acid Glycosyl Component: A UNIQUE O-METHYLATED GLYCAN OF UNIFORM SIZE, CONTAINING 6-DEOXY-3-O-METHYL-D-TALOSE, N-ACETYLQUINOVOSAMINE, AND RHIZOAMINURONIC ACID (3-ACETIMIDOYLAMINO-3-DEOXY-D-GLUCO-HEXURONIC ACID) J. Biol. Chem., June 6, 2008; 283(23): 16037 - 16050. [Abstract] [Full Text] [PDF]

				W. D'Haeze, C. Leoff, G. Freshour, K. D. Noel, and R. W. Carlson Rhizobium etli CE3 Bacteroid Lipopolysaccharides Are Structurally Similar but Not Identical to Those Produced by Cultured CE3 Bacteria J. Biol. Chem., June 8, 2007; 282(23): 17101 - 17113. [Abstract] [Full Text] [PDF]

				A. J.-L. Le Quere, W. J. Deakin, C. Schmeisser, R. W. Carlson, W. R. Streit, W. J. Broughton, and L. S. Forsberg Structural Characterization of a K-antigen Capsular Polysaccharide Essential for Normal Symbiotic Infection in Rhizobium sp. NGR234: DELETION OF THE rkpMNO LOCUS PREVENTS SYNTHESIS OF 5,7-DIACETAMIDO-3,5,7,9-TETRADEOXY-NON-2-ULOSONIC ACID J. Biol. Chem., September 29, 2006; 281(39): 28981 - 28992. [Abstract] [Full Text] [PDF]

				B. L. Reuhs, B. Relic, L. S. Forsberg, C. Marie, T. Ojanen-Reuhs, S. B. Stephens, C.-H. Wong, S. Jabbouri, and W. J. Broughton Structural Characterization of a Flavonoid-Inducible Pseudomonas aeruginosa A-Band-Like O Antigen of Rhizobium sp. Strain NGR234, Required for the Formation of Nitrogen-Fixing Nodules J. Bacteriol., September 15, 2005; 187(18): 6479 - 6487. [Abstract] [Full Text] [PDF]

				K. D. Noel, J. M. Box, and V. J. Bonne 2-O-Methylation of Fucosyl Residues of a Rhizobial Lipopolysaccharide Is Increased in Response to Host Exudate and Is Eliminated in a Symbiotically Defective Mutant Appl. Envir. Microbiol., March 1, 2004; 70(3): 1537 - 1544. [Abstract] [Full Text] [PDF]