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Identification of a novel α(1→6) mannopyranosyltransferase MptB from Corynebacterium glutamicum by deletion of a conserved gene, NCgl1505, affords a lipomannan- and lipoarabinomannan-deficient mutant

By Arun K Mishra, Luke J Alderwick, Doris Rittmann, Cindy Wang, Apoorva Bhatt, William R Jacobs, Kuni Takayama, Lothar Eggeling and Gurdyal S Besra

Abstract

Mycobacterium tuberculosis and Corynebacterium glutamicum share a similar cell wall structure and orthologous enzymes involved in cell wall assembly. Herein, we have studied C. glutamicum NCgl1505, the orthologue of putative glycosyltransferases Rv1459c from M. tuberculosis and MSMEG3120 from Mycobacterium smegmatis. Deletion of NCgl1505 resulted in the absence of lipomannan (Cg-LM-A), lipoarabinomannan (Cg-LAM) and a multi-mannosylated polymer (Cg-LM-B) based on a 1,2-di-O-C16/C18:1-(α-D-glucopyranosyluronic acid)-(1→3)-glycerol (GlcAGroAc2) anchor, while syntheses of triacylated-phosphatidyl-myo-inositol dimannoside (Ac1PIM2) and Man1GlcAGroAc2 were still abundant in whole cells. Cell-free incubation of C. glutamicum membranes with GDP-[14C]Man established that C. glutamicum synthesized a novel α(1→6)-linked linear form of Cg-LM-A and Cg-LM-B from Ac1PIM2 and Man1GlcAGroAc2 respectively. Furthermore, deletion of NCgl1505 also led to the absence of in vitro synthesized linear Cg-LM-A and Cg-LM-B, demonstrating that NCgl1505 was involved in core α(1→6) mannan biosynthesis of Cg-LM-A and Cg-LM-B, extending Ac1PI[14C]M2 and [14C]Man1GlcAGroAc2 primers respectively. Use of the acceptor α-D-Manp-(1→6)-α-D-Manp-O-C8 in an in vitro cell-free assay confirmed NCgl1505 as an α(1→6) mannopyranosyltransferase, now termed MptB. While Rv1459c and MSMEG3120 demonstrated similar in vitroα(1→6) mannopyranosyltransferase activity, deletion of the Rv1459c homologue in M. smegmatis did not result in loss of mycobacterial LM/LAM, indicating a functional redundancy for this enzyme in mycobacteria

Topics: Research Articles
Publisher: Blackwell Publishing Ltd
OAI identifier: oai:pubmedcentral.nih.gov:2440535
Provided by: PubMed Central
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    Citations

    1. (1992). A new subfamily of bacterial ABC-type transport systems catalyzing export of drugs and carbohydrates.
    2. (2004). Acyl-CoA carboxylases (accD2 and accD3), together with a unique polyketide synthase (Cg-pks), are key to mycolic acid biosynthesis in Corynebacterianeae such as Corynebacterium glutamicum and Mycobacterium tuberculosis.
    3. (2008). Analysis of a new mannosyltransferase required for the synthesis of phosphotidylinositol mannosides and lipoarabinomannan reveals two lipomannan pools in Corynebacterineae.
    4. (2006). Arabinan-deficient mutants of Corynebacterium glutamicum and the consequent flux in decaprenylmonophosphoryl-D-arabinose metabolism.
    5. (2005). ATP hydrolysis is required to reset the ATP-binding Identification of a novel a(1,6) mannopyranosyltransferase 1611 ©
    6. (1994). Binding of the terminal mannosyl units of lipoarabinomannan from a virulent strain of Mycobacterium tuberculosis to human macrophages.
    7. (1968). Biosynthesis of mannophosphoinositides by Mycobacterium phlei. Enzymatic acylation of the dimannophosphoinositides.
    8. (1967). Biosynthesis of mannophosphoinositides by Mycobacterium phlei. The family of dimannophosphoinositides.
    9. (1966). Biosynthesis of mannophospholipids by Mycobacterium phlei.
    10. (2006). Biosynthesis of mycobacterial lipoarabinomannan: role of a branching mannosyltransferase.
    11. (1997). Biosynthesis of mycobacterial lipoarabinomannan.
    12. (2004). Biosynthesis of mycobacterial phosphatidylinositol mannosides.
    13. (2002). Characterization of a putative a-mannosyltransferase involved in phosphatidylinositol trimannoside biosynthesis in Mycobacterium tuberculosis.
    14. (1997). Clinical microbiology of coryneform bacteria.
    15. (2004). Comparative cell wall core biosynthesis in the mycolated pathogens, Mycobacterium tuberculosis and Corynebacterium diphtheriae.
    16. (1990). Coryneform bacteria in infectious diseases: clinical and laboratory aspects.
    17. (2002). Definition of the first mannosylation step in phosphatidylinositol mannoside synthesis. PimA is essential for growth of mycobacteria.
    18. (2005). Deletion of Cg-emb in corynebacterianeae leads to a novel truncated cell wall arabinogalactan, whereas inactivation of Cg-ubiA results in an arabinan-deficient mutant with a cell wall galactan core.
    19. (2004). DIALIGN: multiple DNA and protein sequence alignment at BiBiServ.
    20. (2003). Disruption of Cg-Ppm1, a polyprenyl monophosphomannose synthase, and the generation of lipoglycan-less mutants in Corynebacterium glutamicum.
    21. (2002). Evidence for a partial redundancy of the fibronectin-binding proteins for the transfer of mycoloyl residues onto the cell wall arabinogalactan termini of Mycobacterium tuberculosis.
    22. (1990). Evidence for the nature of the link between the arabinogalactan and peptidoglycan of mycobacterial cell walls.
    23. (2001). Galactan biosynthesis in Mycobacterium tuberculosis. Identification of a bifunctional UDP-galactofuranosyltransferase.
    24. (2006). Genetic basis for the synthesis of the immunomodulatory mannose caps of lipoarabinomannan in Mycobacterium tuberculosis.
    25. (2005). Handbook of Corynebacterium Glutamicum.
    26. (1997). Hydrophobic mannosides act as acceptors for trypanosome a-mannosyltransferases.
    27. (2006). Identification of a Novel arabinofuranosyltransferase (AftA) involved in cell wall arabinan biosynthesis in Mycobacterium tuberculosis.
    28. (2007). Identification of a novel arabinofuranosyltransferase AftB involved in a terminal step of cell wall arabinan biosynthesis in Corynebacterianeae, such as Corynebacterium glutamicum and Mycobacterium tuberculosis.
    29. (2007). Identification of an a(1–6) mannopyranosyltransferase (MptA), involved in Corynebacterium glutamicum lipomanann biosynthesis, and identification of its orthologue in Mycobacterium tuberculosis.
    30. (2003). Identification of the required acyltransferase step in the biosynthesis of the phosphatidylinositol mannosides of Mycobacterium species.
    31. (2007). Inactivation of Corynebacterium glutamicum NCgl0452 and the role of MgtA in the biosynthesis of a novel mannosylated glycolipid involved in lipomannan biosynthesis.
    32. (1995). Inositol phosphate capping of the nonreducing termini of lipoarabinomannan from rapidly growing strains of Mycobacterium.
    33. (1998). Lipoarabinomannan of Mycobacterium tuberculosis promotes protein tyrosine dephosphorylation and inhibition of mitogen-activated protein kinase in human mononuclear phagocytes. Role of the Src homology 2 containing tyrosine phosphatase 1.
    34. (2003). Lipoarabinomannans: from structure to biosynthesis.
    35. (1991). Location of the mycolyl ester substituents in the cell walls of mycobacteria.
    36. (1993). Major struc1610 A. K. Mishra etal. ©
    37. (2008). Mishra etal. ©
    38. (1998). Mycobacterial lipoarabinomannan: an extraordinary lipoheteroglycan with profound physiological effects.
    39. (2002). Mycobacterial lipoarabinomannans: modulators of dendritic cell function and the apoptotic response.
    40. (1997). Mycobacterium smegmatis phosphoinositols-glyceroarabinomannans. Structure and localization of alkali-labile and alkali-stable phosphoinositides.
    41. (2003). Mycobacterium tuberculosis glycosylated phosphatidylinositol causes phagosome maturation arrest.
    42. (2002). Mycobacterium tuberculosis phoP mutant: lipoarabinomannan molecular structure.
    43. (2000). Mycobacterium tuberculosis-activated dendritic cells induce protective immunity in mice.
    44. (2007). New insights into the biosynthesis of mycobacterial lipomannan arising from deletion of a conserved gene.
    45. (2006). PimE Is a Polyprenolphosphate-mannose-dependent mannosyltransferase that transfers the fifth mannose of phosphatidylinositol mannoside in mycobacteria.
    46. (2002). Ppm1, a novel polyprenol monophosphomannose synthase from Mycobacterium tuberculosis.
    47. (1990). Predominant structural features of the cell wall arabinogalactan of Mycobacterium tuberculosis as revealed through characterization of oligoglycosyl alditol fragments by gas chromatography/mass spectrometry and by 1H and 13C NMR analyses.
    48. (1997). Proposal for a new hierarchic classification system, Actinobacteria classis nov.
    49. (2006). Protein splicing of SufB is crucial for the functionality of the Mycobacterium tuberculosis SUF machinery.
    50. (1993). Regulation of murine macrophage effector functions by lipoarabinomannan from mycobacterial strains with different degrees of virulence.
    51. (2005). Roles of conserved proline and glycosyltransferase motifs of EmbC in biosynthesis of lipoarabinomannan.
    52. (1998). Sequence analyses and phylogenetic characterization of the ZIP family of metal ion transport proteins.
    53. (1994). Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum.
    54. (2002). Specialized transduction: an efficient method for generating marked and unmarked targeted gene disruptions
    55. (2008). Structural characterization and functional properties of a novel lipomannan variant isolated from a Corynebacterium glutamicum pimB mutant. Antonie Van Leeuwenhoek
    56. (1999). Structural definition of arabinomannans from Mycobacterium bovis BCG.
    57. (1993). Structural definition of the non-reducing termini of mannose-capped LAM from Mycobacterium tuberculosis through selective enzymatic degradation and fast atom bombardment-mass spectrometry.
    58. (2002). Structural study of lipomannan and lipoarabinomannan from Mycobacterium chelonae. Presence of unusual components with a1,3-mannopyranose side chains.
    59. (1986). Structure and antigenicity of the phosphorylated lipopolysaccharide antigens from the leprosy and tubercle bacilli.
    60. (2003). Structure, function, and biogenesis of the cell wall of Mycobacterium tuberculosis.
    61. (2001). Synthetic mannosides act as acceptors for mycobacterial a1–6 mannosyltransferase.
    62. (1985). Systematic analysis of complex mycobacterial lipids.
    63. (2003). The cell surface receptor DC-SIGN discriminates between Mycobacterium species through selective recognition of the mannose caps on lipoarabinomannan.
    64. (2003). The Emb proteins of mycobacteria direct arabinosylation of lipoarabinomannan and arabinogalactan via an N-terminal recognition region and a C-terminal synthetic region.
    65. (1996). The embAB genes of Mycobacterium avium encode an arabinosyl transferase involved in cell wall arabinan biosynthesis that is the target for the antimycobacterial drug ethambutol.
    66. (1995). The envelope of mycobacteria.
    67. (2007). The Glycosyltransferases of Mycobacterium tuberculosis; roles in the synthesis of arabinogalactan, lipoarabinomannan, and other glycoconjugates.
    68. (2005). The HHpred interactive server for protein homology detection and structure prediction.
    69. (1999). The LysE superfamily: topology of the lysine exporter LysE of Corynebacterium glutamicum, a paradyme for a novel superfamily of transmembrane solute translocators.
    70. (2007). The mannose cap of mycobacterial lipoarabinomannan does not dominate the Mycobacterium–host interaction.
    71. (2002). The methyl-branched fortifications of Mycobacterium tuberculosis.
    72. (1997). The mycobacterial cell wall: biosynthesis of arabinogalactan and lipoarabinomannan.
    73. (1997). The phosphatidylmyo-inositol anchor of the lipoarabinomannans from Mycobacterium bovis bacillus Calmette Guerin. Heterogeneity, structure, and role in the regulation of cytokine secretion.
    74. (1999). The pimB gene of Mycobacterium tuberculosis encodes a mannosyltransferase involved in lipoarabinomannan biosynthesis.
    75. (2003). Three monophyletic superfamilies account for the majority of the known glycosyltransferases.
    76. (2006). Transposome mutagenesis of an integral membrane transporter in Corynebacterium matruchotii.
    77. (1992). Tuberculosis: commentary on a reemergent killer.

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