Abstract

This revised application is to support the investigation of the structure and biosynthesis of the polysaccharide components of the LPS molecules involved in Rhyzobial symbiosis. These cell wall polysaccharides are thought to be important determinants of virulence and to be required for the invasion of the host. The overall goal is to determine the structure and biosynthesis of these complex glycolipids and determine the specific structural features required for their biological activity.
The specific aims are: 1) characterize the minimum lipidA core structure requirements for the ligation of the O-chain polysaccharide, and characterize the modification to this minimal lipid. A core structure which lead to the ligation of increased O-chain polysaccharide, 2) to characterize the structural modification to the O-chain that occur during the symbiotic infections, 3) to isolate mutants of R. leguminosarum that are unable to add unique structural features normal found in the LPS of this species, and 4) to characterize the R. leguminosarum specific enzymes that process the Kdo2-lipid-IVA precursor to the unique structure of this organism.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM039583-13
Application #
6179566
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Program Officer
Chin, Jean
Project Start
1988-06-01
Project End
2002-03-31
Budget Start
2000-04-01
Budget End
2001-03-31
Support Year
13
Fiscal Year
2000
Total Cost
$199,534
Indirect Cost

  Institution

Name
University of Georgia
Department
Type
Organized Research Units
DUNS #
City
Athens
State
GA
Country
United States
Zip Code
30602

  Publications

Bourassa, Dianna V; Kannenberg, Elmar L; Sherrier, D Janine et al. (2017) The Lipopolysaccharide Lipid A Long-Chain Fatty Acid Is Important for Rhizobium leguminosarum Growth and Stress Adaptation in Free-Living and Nodule Environments. Mol Plant Microbe Interact 30:161-175
Brown, Dusty B; Muszynski, Artur; Carlson, Russell W (2013) Elucidation of a novel lipid A ?-(1,1)-GalA transferase gene (rgtF) from Mesorhizobium loti: Heterologous expression of rgtF causes Rhizobium etli to synthesize lipid A with ?-(1,1)-GalA. Glycobiology 23:546-58
Brown, Dusty B; Muszynski, Artur; Salas, Omar et al. (2013) Elucidation of the 3-O-deacylase gene, pagL, required for the removal of primary ?-hydroxy fatty acid from the lipid A in the nitrogen-fixing endosymbiont Rhizobium etli CE3. J Biol Chem 288:12004-13
Brown, Dusty B; Forsberg, L Scott; Kannenberg, Elmar L et al. (2012) Characterization of galacturonosyl transferase genes rgtA, rgtB, rgtC, rgtD, and rgtE responsible for lipopolysaccharide synthesis in nitrogen-fixing endosymbiont Rhizobium leguminosarum: lipopolysaccharide core and lipid galacturonosyl residues confer me J Biol Chem 287:935-49
Muszynski, Artur; Laus, Marc; Kijne, Jan W et al. (2011) Structures of the lipopolysaccharides from Rhizobium leguminosarum RBL5523 and its UDP-glucose dehydrogenase mutant (exo5). Glycobiology 21:55-68
Brown, Dusty B; Huang, Yu-Chu; Kannenberg, Elmar L et al. (2011) An acpXL mutant of Rhizobium leguminosarum bv. phaseoli lacks 27-hydroxyoctacosanoic acid in its lipid A and is developmentally delayed during symbiotic infection of the determinate nodulating host plant Phaseolus vulgaris. J Bacteriol 193:4766-78
Vanderlinde, Elizabeth M; Harrison, Joe J; Muszy?ski, Artur et al. (2010) Identification of a novel ABC transporter required for desiccation tolerance, and biofilm formation in Rhizobium leguminosarum bv. viciae 3841. FEMS Microbiol Ecol 71:327-40
Vanderlinde, Elizabeth M; Muszynski, Artur; Harrison, Joe J et al. (2009) Rhizobium leguminosarum biovar viciae 3841, deficient in 27-hydroxyoctacosanoate-modified lipopolysaccharide, is impaired in desiccation tolerance, biofilm formation and motility. Microbiology 155:3055-69
Forsberg, L Scott; Carlson, Russell W (2008) 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 s J Biol Chem 283:16037-50
D'Haeze, Wim; Leoff, Christine; Freshour, Glenn et al. (2007) Rhizobium etli CE3 bacteroid lipopolysaccharides are structurally similar but not identical to those produced by cultured CE3 bacteria. J Biol Chem 282:17101-13

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