title: MKAN27435 is required for the biosynthesis of higher subclasses of lipooligosaccharides in Mycobacterium kansasii.
abstract: Lipooligosaccharides are glycolipids found in the cell wall of many mycobacterial species including the opportunistic pathogen Mycobacterium kansasii. The genome of M. kansasii ATCC12478 contains a cluster with genes orthologous to Mycobacterium marinum LOS biosynthesis genes. To initiate a genetic dissection of this cluster and demonstrate its role in LOS biosynthesis in M. kansasii, we chose MKAN27435, a gene encoding a putative glycosyltransferase. Using Specialized Transduction, a phage-based gene knockout tool previously used to generate null mutants in other mycobacteria, we generated a MKAN27435 null mutant. The mutant strain was found to be defective in the biosynthesis of higher LOS subspecies, viz LOS-IV, LOS-V, LOS-VI and LOS-VII. Additionally, a range of low abundance species were detected in the mutant strain and mass spectroscopic analysis indicated that these were shunt products generated from LOS-III by the addition of up to six molecules of a pentose.
authors.affiliation: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom.
authors.forename: Vijayashankar
authors.initials: V
authors.lastname: Nataraj
authors.affiliation: Department of Life Sciences, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Poh-choo
authors.initials: PC
authors.lastname: Pang
authors.affiliation: Department of Life Sciences, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Stuart M
authors.initials: SM
authors.lastname: Haslam
authors.affiliation: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom.
authors.forename: Natacha
authors.initials: N
authors.lastname: Veerapen
authors.affiliation: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom.
authors.forename: David E
authors.initials: DE
authors.lastname: Minnikin
authors.affiliation: Department of Life Sciences, Imperial College London, London, SW7 2AZ, United Kingdom.
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.affiliation: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom.
authors.forename: Gurdyal S
authors.initials: GS
authors.lastname: Besra
authors.affiliation: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom.
authors.forename: Apoorva
authors.initials: A
authors.lastname: Bhatt
mesh_terms: D055786:Gene Knockout Techniques; D016695:Glycosyltransferases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry; D005810:Multigene Family / Q000235:genetics; D009154:Mutation; D019909:Mycobacterium kansasii / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D016366:Open Reading Frames / Q000235:genetics
chemical_list: D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; D016695:Glycosyltransferases