| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Genetics and Molecular Biology |
Department of Microbiology, University of Guelph, Guelph, Ontario , Canada N1G 2W11
Author for correspondence: Joseph S. Lam. Tel: +1 519 824 4120 Ext. 3823. Fax: +1 519 837 1802. e-mail: jlam{at}uoguelph.ca
This study reports the organization of the wbp gene cluster and characterization of a number of genes that are essential for B-band O antigen biosynthesis in the clinically prevalent Pseudomonas aeruginosa serotype O6. Twelve genes were identified that share homology with other LPS and polysaccharide biosynthetic genes. This cluster contains homologues of wzx (encoding the O antigen flippase/translocase) and wzz (which modulates O antigen chain length distribution) genes, typical of a wzy- dependent pathway. However, a complete wzy gene (encoding the O-polymerase) was not found within the cluster. Four biosynthetic genes, wbpO, wbpP, wbpV and wbpM, and four putative glycosyltransferase genes, wbpR, wbpT , wbpU and wbpL, were identified in the cluster. To characterize their roles in LPS biosynthesis, null mutants of wbpO, wbpP, wbpV, wbpL and wbpM were generated using a gene-replacement strategy. Mutations in each of these genes caused deficiency in B-band synthesis. The wbpL mutant was deficient in both A-band and B-band LPS. WbpLO6 is a bi-functional enzyme which could initiate B-band synthesis through the addition of QuiNAc to undecaprenol phosphate, and A-band synthesis by transferring either a GalNAc or a GlcNAc residue. Another approach used to assign function to the wbpO6 genes was by complementation analysis. Two genes from Salmonella typhi, wcdA and wcdB, responsible for the synthesis of a homopolymer of GalNAcA called Vi antigen were used in complementation experiments to verify the functions of wbpO and wbpP. wcdA and wcdB restored B-band synthesis in wbpO and wbpP mutants respectively, implying that wbpO and wbpP are involved in UDP-GalNAcA synthesis. Although wbpV has homology to wbpK of the serotype O5 B-band LPS synthesis cluster, complementation analysis using the respective null mutants showed that the genes are not interchangeable. A knockout mutation of wbpN (located downstream of wbpM) did not abrogate LPS synthesis in either O5 or O6; therefore, it has been renamed orf48.5. These results establish the organization of genes involved in P. aeruginosa B-band O antigen synthesis and provide the evidence to assign functions to a number of LPS biosynthetic genes.
Keywords: Pseudomonas aeruginosa, lipopolysaccharide, rfb, wbp, O antigen
Abbreviations: GlcNAc, N-acetylglucosamine; GalNAc, N- acetylgalactosamine; GalNAcA, 2-deoxy-2-N- acetylgalactosaminuronic acid; GalNAcAN, 2-acetamido-2-deoxy-D- galacturonic acid (N-acetyl-D-galactosaminuronic acid); GalNFmA, 2-deoxy-2-formamido-D-galacturonic acid (N- formylgalactosaminuronic acid); QuiNAc, 2-acetamido-2,6- dideoxy-D-glucose (N-acetylquinovosamine); Rha, rhamnose; Fuc2NAc, 2-acetamido-2,6-dideoxy-D-galactose (6- deoxy-N-acetylgalactosamine)
The GenBank accession number for the sequence reported in this paper is AF035937.
a Present address: Center for Infection and Biomaterials Research, Toronto General Hospital, 200 Elizabeth St, Toronto, ON, Canada M5G 2C4.
This article has been cited by other articles:
|
|
 |
|
  E. L. Westman, A. Preston, R. A. Field, and J. S. Lam Biosynthesis of a Rare Di-N-Acetylated Sugar in the Lipopolysaccharides of both Pseudomonas aeruginosa and Bordetella pertussis Occurs via an Identical Scheme despite Different Gene Clusters J. Bacteriol., September 15, 2008; 190(18): 6060 - 6069. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. L. Miller, M. J. Matewish, D. J. McNally, N. Ishiyama, E. M. Anderson, D. Brewer, J.-R. Brisson, A. M. Berghuis, and J. S. Lam Flagellin Glycosylation in Pseudomonas aeruginosa PAK Requires the O-antigen Biosynthesis Enzyme WbpO J. Biol. Chem., February 8, 2008; 283(6): 3507 - 3518. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. M. Cunneen and P. R. Reeves The Yersinia kristensenii O11 O-Antigen Gene Cluster was Acquired by Lateral Gene Transfer and Incorporated at a Novel Chromosomal Locus Mol. Biol. Evol., June 1, 2007; 24(6): 1355 - 1365. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Sebastian, S. T. Dillon, J. G. Lynch, L. T. Blalock, E. Balon, K. T. Lee, L. E. Comstock, J. W. Conlan, E. J. Rubin, A. O. Tzianabos, et al. A Defined O-Antigen Polysaccharide Mutant of Francisella tularensis Live Vaccine Strain Has Attenuated Virulence while Retaining Its Protective Capacity Infect. Immun., May 1, 2007; 75(5): 2591 - 2602. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Raynaud, K. L. Meibom, M.-A. Lety, I. Dubail, T. Candela, E. Frapy, and A. Charbit Role of the wbt Locus of Francisella tularensis in Lipopolysaccharide O-Antigen Biogenesis and Pathogenicity Infect. Immun., January 1, 2007; 75(1): 536 - 541. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. A. Knirel, O. V. Bystrova, N. A. Kocharova, U. Zahringer, and G. B. Pier Review: Conserved and variable structural features in the lipopolysaccharide of Pseudomonas aeruginosa Innate Immunity, December 1, 2006; 12(6): 324 - 336. [Abstract] [PDF]
|
|
|
|
 |
|
 J. M. Struble and R. T. Gill Reverse Engineering Antibiotic Sensitivity in a Multidrug-Resistant Pseudomonas aeruginosa Isolate. Antimicrob. Agents Chemother., July 1, 2006; 50(7): 2506 - 2515. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Y. Park, J. H. Lee, M. W. Kim, H. G. Jeong, B. C. Lee, T. S. Kim, and S. H. Choi Identification of the Vibrio vulnificus wbpP Gene and Evaluation of Its Role in Virulence Infect. Immun., January 1, 2006; 74(1): 721 - 728. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. P. Price and F. A. Momany Modeling bacterial UDP-HexNAc: polyprenol-P HexNAc-1-P transferases Glycobiology, September 1, 2005; 15(9): 29R - 42R. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Ishiyama, C. Creuzenet, J. S. Lam, and A. M. Berghuis Crystal Structure of WbpP, a Genuine UDP-N-acetylglucosamine 4-Epimerase from Pseudomonas aeruginosa: SUBSTRATE SPECIFICITY IN UDP-HEXOSE 4-EPIMERASES J. Biol. Chem., May 21, 2004; 279(21): 22635 - 22642. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. S. Forsberg, K. D. Noel, J. Box, and R. W. Carlson 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 J. Biol. Chem., December 19, 2003; 278(51): 51347 - 51359. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. L. Prior, R. G. Prior, P. G. Hitchen, H. Diaper, K. F. Griffin, H. R. Morris, A. Dell, and R. W. Titball Characterization of the O antigen gene cluster and structural analysis of the O antigen of Francisella tularensis subsp. tularensis J. Med. Microbiol., October 1, 2003; 52(10): 845 - 851. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. Lawrence, M. M. Banes, P. Azadi, and B. Y. Reeks The Edwardsiella ictaluri O polysaccharide biosynthesis gene cluster and the role of O polysaccharide in resistance to normal catfish serum and catfish neutrophils Microbiology, June 1, 2003; 149(6): 1409 - 1421. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Kapatral, N. Ivanova, I. Anderson, G. Reznik, A. Bhattacharyya, W. L. Gardner, N. Mikhailova, A. Lapidus, N. Larsen, M. D'Souza, et al. Genome Analysis of F. nucleatum sub spp vincentii and Its Comparison With the Genome of F. nucleatum ATCC 25586 Genome Res., June 1, 2003; 13(6): 1180 - 1189. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. H. Spencer, A. Kas, E. E. Smith, C. K. Raymond, E. H. Sims, M. Hastings, J. L. Burns, R. Kaul, and M. V. Olson Whole-Genome Sequence Variation among Multiple Isolates of Pseudomonas aeruginosa J. Bacteriol., February 15, 2003; 185(4): 1316 - 1325. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. N. Parsons, S. Panagea, C. H. M. Smart, M. J. Walshaw, C. A. Hart, and C. Winstanley Use of Subtractive Hybridization To Identify a Diagnostic Probe for a Cystic Fibrosis Epidemic Strain of Pseudomonas aeruginosa J. Clin. Microbiol., December 1, 2002; 40(12): 4607 - 4611. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D.-Q. Xu, J. O. Cisar, N. Ambulos Jr., D. H. Burr, and D. J. Kopecko Molecular Cloning and Characterization of Genes for Shigella sonnei Form I O Polysaccharide: Proposed Biosynthetic Pathway and Stable Expression in a Live Salmonella Vaccine Vector Infect. Immun., August 1, 2002; 70(8): 4414 - 4423. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Creuzenet, R. V. Urbanic, and J. S. Lam Structure-Function Studies of Two Novel UDP-GlcNAc C6 Dehydratases/C4 Reductases. VARIATION FROM THE SYK DOGMA J. Biol. Chem., July 19, 2002; 277(30): 26769 - 26778. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. K. Raymond, E. H. Sims, A. Kas, D. H. Spencer, T. V. Kutyavin, R. G. Ivey, Y. Zhou, R. Kaul, J. B. Clendenning, and M. V. Olson Genetic Variation at the O-Antigen Biosynthetic Locus in Pseudomonas aeruginosa J. Bacteriol., July 1, 2002; 184(13): 3614 - 3622. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Suzuki, T. Kanagawa, and Y. Kamagata Identification of a Gene Essential for Sheathed Structure Formation in Sphaerotilus natans, a Filamentous Sheathed Bacterium Appl. Envir. Microbiol., January 1, 2002; 68(1): 365 - 371. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. R. Dean, A. Datta, R. W. Carlson, and J. B. Goldberg WbjA Adds Glucose To Complete the O-Antigen Trisaccharide Repeating Unit of the Lipopolysaccharide of Pseudomonas aeruginosa Serogroup O11 J. Bacteriol., January 1, 2002; 184(1): 323 - 326. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Rahim, L. L. Burrows, M. A. Monteiro, M. B. Perry, and J. S. Lam Involvement of the rml locus in core oligosaccharide and O polysaccharide assembly in Pseudomonas aeruginosa Microbiology, November 1, 2000; 146(11): 2803 - 2814. [Abstract] [Full Text]
|
|
|
|
|
|
K. D. Noel, L. S. Forsberg, and R. W. Carlson Varying the Abundance of O Antigen in Rhizobium etli and Its Effect on Symbiosis with Phaseolus vulgaris J. Bacteriol., October 1, 2000; 182(19): 5317 - 5324. [Abstract] [Full Text]
|
|
|
|
|
|
H. Yang, M. Matewish, I. Loubens, D. G. Storey, J. S. Lam, and S. Jin migA, a quorum-responsive gene of Pseudomonas aeruginosa, is highly expressed in the cystic fibrosis lung environment and modifies low-molecular-mass lipopolysaccharide Microbiology, October 1, 2000; 146(10): 2509 - 2519. [Abstract] [Full Text]
|
|
|
|
|
|
L. L. Burrows, R. V. Urbanic, and J. S. Lam Functional Conservation of the Polysaccharide Biosynthetic Protein WbpM and Its Homologues in Pseudomonas aeruginosa and Other Medically Significant Bacteria Infect. Immun., February 1, 2000; 68(2): 931 - 936. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Creuzenet, M. Belanger, W. W. Wakarchuk, and J. S. Lam Expression, Purification, and Biochemical Characterization of WbpP, a New UDP-GlcNAc C4 Epimerase from Pseudomonas aeruginosa Serotype O6 J. Biol. Chem., June 16, 2000; 275(25): 19060 - 19067. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Zhao, C. Creuzenet, M. Belanger, E. Egbosimba, J. Li, and J. S. Lam WbpO, a UDP-N-acetyl-D-galactosamine Dehydrogenase from Pseudomonas aeruginosa Serotype O6 J. Biol. Chem., October 20, 2000; 275(43): 33252 - 33259. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
