A Bifunctional Enzyme in a Single Gene Catalyzes the Incorporation of GlcN into the Aeromonas Core Lipopolysaccharide

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2009 Oct 7

PMID 19805547

Citations 9

Authors

Natalia Jimenez

Silvia Vilches

Anna Lacasta

Miguel Regue

Susana Merino

Juan M Tomas

Affiliations

Soon will be listed here.

Abstract

The core lipopolysaccharide (LPS) of Aeromonas hydrophila AH-3 and Aeromonas salmonicida A450 is characterized by the presence of the pentasaccharide alpha-d-GlcN-(1-->7)-l-alpha-d-Hep-(1-->2)-l-alpha-d-Hep-(1-->3)-l-alpha-d-Hep-(1-->5)-alpha-Kdo. Previously it has been suggested that the WahA protein is involved in the incorporation of GlcN residue to outer core LPS. The WahA protein contains two domains: a glycosyltransferase and a carbohydrate esterase. In this work we demonstrate that the independent expression of the WahA glycosyltransferase domain catalyzes the incorporation of GlcNAc from UDP-GlcNAc to the outer core LPS. Independent expression of the carbohydrate esterase domain leads to the deacetylation of the GlcNAc residue to GlcN. Thus, the WahA is the first described bifunctional glycosyltransferase enzyme involved in the biosynthesis of core LPS. By contrast in Enterobacteriaceae containing GlcN in their outer core LPS the two reactions are performed by two different enzymes.

Citing Articles

Bifunctional and monofunctional α-neoagarooligosaccharide hydrolases from Streptomyces coelicolor A3(2).

Tsevelkhoroloo M, Dhakshnamoorthy V, Hong Y, Lee C, Hong S Appl Microbiol Biotechnol. 2023; 107(12):3997-4008.

PMID: 37184654 DOI: 10.1007/s00253-023-12552-x.

Comparative Genomics of the Aeromonadaceae Core Oligosaccharide Biosynthetic Regions.

Forn-Cuni G, Merino S, Tomas J Int J Mol Sci. 2017; 18(3).

PMID: 28264491 PMC: 5372535. DOI: 10.3390/ijms18030519.

The FlgT Protein Is Involved in Aeromonas hydrophila Polar Flagella Stability and Not Affects Anchorage of Lateral Flagella.

Merino S, Tomas J Front Microbiol. 2016; 7:1150.

PMID: 27507965 PMC: 4960245. DOI: 10.3389/fmicb.2016.01150.

Polar Glycosylated and Lateral Non-Glycosylated Flagella from Aeromonas hydrophila Strain AH-1 (Serotype O11).

Fulton K, Mendoza-Barbera E, Twine S, Tomas J, Merino S Int J Mol Sci. 2015; 16(12):28255-69.

PMID: 26633358 PMC: 4691044. DOI: 10.3390/ijms161226097.

The polar and lateral flagella from Plesiomonas shigelloides are glycosylated with legionaminic acid.

Merino S, Aquilini E, Fulton K, Twine S, Tomas J Front Microbiol. 2015; 6:649.

PMID: 26167161 PMC: 4481668. DOI: 10.3389/fmicb.2015.00649.

References

Reith M, Singh R, Curtis B, Boyd J, Bouevitch A, Kimball J . The genome of Aeromonas salmonicida subsp. salmonicida A449: insights into the evolution of a fish pathogen. BMC Genomics. 2008; 9:427. PMC: 2556355. DOI: 10.1186/1471-2164-9-427. View

Merino S, Camprubi S, Tomas J . The role of lipopolysaccharide in complement-killing of Aeromonas hydrophila strains of serotype O:34. J Gen Microbiol. 1991; 137(7):1583-90. DOI: 10.1099/00221287-137-7-1583. View

Boyer H . A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969; 41(3):459-72. DOI: 10.1016/0022-2836(69)90288-5. View

Scott M . The pathogenicity of Aeromonas salmonicida (Griffin) in sea and brackish waters. J Gen Microbiol. 1968; 50(2):321-7. DOI: 10.1099/00221287-50-2-321. View

Casadaban M, Cohen S . Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980; 138(2):179-207. DOI: 10.1016/0022-2836(80)90283-1. View

Darveau R, Hancock R . Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J Bacteriol. 1983; 155(2):831-8. PMC: 217756. DOI: 10.1128/jb.155.2.831-838.1983. View

Valvano M, Marolda C, Bittner M, Simon T, Klena J . The rfaE gene from Escherichia coli encodes a bifunctional protein involved in biosynthesis of the lipopolysaccharide core precursor ADP-L-glycero-D-manno-heptose. J Bacteriol. 2000; 182(2):488-97. PMC: 94300. DOI: 10.1128/JB.182.2.488-497.2000. View

Vinogradov E, Bock K, Holst O, Brade H . The structure of the lipid A-core region of the lipopolysaccharides from Vibrio cholerae O1 smooth strain 569B (Inaba) and rough mutant strain 95R (Ogawa). Eur J Biochem. 1995; 233(1):152-8. DOI: 10.1111/j.1432-1033.1995.152_1.x. View

Lairson L, Henrissat B, Davies G, Withers S . Glycosyltransferases: structures, functions, and mechanisms. Annu Rev Biochem. 2008; 77:521-55. DOI: 10.1146/annurev.biochem.76.061005.092322. View

10.

Wang Z, Li J, Vinogradov E, Altman E . Structural studies of the core region of Aeromonas salmonicida subsp. salmonicida lipopolysaccharide. Carbohydr Res. 2005; 341(1):109-17. DOI: 10.1016/j.carres.2005.10.017. View

11.

Milton D, OToole R, Horstedt P, Wolf-Watz H . Flagellin A is essential for the virulence of Vibrio anguillarum. J Bacteriol. 1996; 178(5):1310-9. PMC: 177804. DOI: 10.1128/jb.178.5.1310-1319.1996. View

12.

McArthur F, Andersson C, Loutet S, Mowbray S, Valvano M . Functional analysis of the glycero-manno-heptose 7-phosphate kinase domain from the bifunctional HldE protein, which is involved in ADP-L-glycero-D-manno-heptose biosynthesis. J Bacteriol. 2005; 187(15):5292-300. PMC: 1196024. DOI: 10.1128/JB.187.15.5292-5300.2005. View

13.

Thompson J, Higgins D, Gibson T . CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994; 22(22):4673-80. PMC: 308517. DOI: 10.1093/nar/22.22.4673. View

14.

Guzman L, Belin D, CARSON M, Beckwith J . Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol. 1995; 177(14):4121-30. PMC: 177145. DOI: 10.1128/jb.177.14.4121-4130.1995. View

15.

Regue M, Izquierdo L, Fresno S, Jimenez N, Pique N, Corsaro M . The incorporation of glucosamine into enterobacterial core lipopolysaccharide: two enzymatic steps are required. J Biol Chem. 2005; 280(44):36648-56. DOI: 10.1074/jbc.M506278200. View

16.

Merino S, Rubires X, Aguilar A, Tomas . The O:34-antigen lipopolysaccharide as an adhesin in Aeromonas hydrophila. FEMS Microbiol Lett. 1996; 139(2-3):97-101. DOI: 10.1111/j.1574-6968.1996.tb08186.x. View

17.

Brozek K, Hosaka K, Robertson A, Raetz C . Biosynthesis of lipopolysaccharide in Escherichia coli. Cytoplasmic enzymes that attach 3-deoxy-D-manno-octulosonic acid to lipid A. J Biol Chem. 1989; 264(12):6956-66. View

18.

Valvano M, Messner P, Kosma P . Novel pathways for biosynthesis of nucleotide-activated glycero-manno-heptose precursors of bacterial glycoproteins and cell surface polysaccharides. Microbiology (Reading). 2002; 148(Pt 7):1979-1989. DOI: 10.1099/00221287-148-7-1979. View

19.

Figurski D, Helinski D . Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979; 76(4):1648-52. PMC: 383447. DOI: 10.1073/pnas.76.4.1648. View

20.

Merino S, Camprubi S, Tomas J . Effect of growth temperature on outer membrane components and virulence of Aeromonas hydrophila strains of serotype O:34. Infect Immun. 1992; 60(10):4343-9. PMC: 257471. DOI: 10.1128/iai.60.10.4343-4349.1992. View