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Nataraj V, Pang PC, Haslam SM, Veerapen N, Minnikin DE, Dell A, Besra GS, Bhatt A
MKAN27435 is required for the biosynthesis of higher subclasses of lipooligosaccharides in Mycobacterium kansasii
PLoS One 10(3) (2015)
ID e0122804
|
LIP-(1-6)-+
|
LIP-(1-3)-+ |
| |
{{{-b-D-Xylp-(1-4)-}}}/n=1-3/-a-L-Rhap-(1-3)-b-D-Glcp-(1-4)-b-D-Glcp-(1-4)-a-D-Glcp-(1-1)-a-D-Glcp
|
LIP-(1-4)-+ |
Show graphically |
Mycobacterium kansasii ATCC 12478
(NCBI TaxID 557599,
species name lookup)
Mycobacterium kansasii ATCC 12478 (ΔMKAN27435 mutant)
(Ancestor NCBI TaxID 557599,
species name lookup)
Taxonomic group: bacteria / Actinobacteria
(Phylum: Actinobacteria)
Associated disease: infection due to Mycobacterium kansasii [ICD11:
XN3L9 
]
NCBI PubMed ID: 25893968Publication DOI: 10.1371/journal.pone.0122804Journal NLM ID: 101285081Publisher: San Francisco, CA: Public Library of Science
Correspondence: a.bhatt
bham.ac.uk
Institutions: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, UK, Department of Life Sciences, Imperial College London, London, UK
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
biosynthesis, Mycobacterium kansasii, lipooligosaccharides (LOS)
Structure type: oligomer
Location inside paper: Fig. 1, A
Compound class: lipooligosaccharide
Contained glycoepitopes: IEDB_114701,IEDB_136105,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_167188,IEDB_174332,IEDB_189517,IEDB_225177,IEDB_742521,IEDB_885823,IEDB_983931,SB_192
Methods: methylation, PCR, DNA techniques, MALDI-TOF MS, radiolabeling, Southern blotting, extraction, cell growth, cloning plasmids, autoradiography, 2D-TLC
Related record ID(s): 49319, 49320
NCBI Taxonomy refs (TaxIDs): 557599
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There is only one chemically distinct structure:
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Nataraj V, Pang PC, Haslam SM, Veerapen N, Minnikin DE, Dell A, Besra GS, Bhatt A
MKAN27435 is required for the biosynthesis of higher subclasses of lipooligosaccharides in Mycobacterium kansasii
PLoS One 10(3) (2015)
ID e0122804
|
LIP-(1-6)-+
|
LIP-(1-3)-+ |
| |
Lac2Me-(1-3)-b-D-QuipNMe-(1-3)-a-L-Fucp-(1-4)-{{{-b-D-Xylp-(1-4)-}}}/n=4-6/-a-L-Rhap2Ac3Me-(1-3)-b-D-Glcp-(1-4)-b-D-Glcp-(1-4)-a-D-Glcp-(1-1)-a-D-Glcp
|
LIP-(1-4)-+ |
Show graphically |
Mycobacterium kansasii ATCC 12478
(NCBI TaxID 557599,
species name lookup)
Taxonomic group: bacteria / Actinobacteria
(Phylum: Actinobacteria)
Associated disease: infection due to Mycobacterium kansasii [ICD11:
XN3L9 ]
NCBI PubMed ID: 25893968Publication DOI: 10.1371/journal.pone.0122804Journal NLM ID: 101285081Publisher: San Francisco, CA: Public Library of Science
Correspondence: a.bhatt
bham.ac.uk
Institutions: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, UK, Department of Life Sciences, Imperial College London, London, UK
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
biosynthesis, Mycobacterium kansasii, lipooligosaccharides (LOS)
Structure type: oligomer
Location inside paper: Fig. 1, B
Compound class: lipooligosaccharide
Contained glycoepitopes: IEDB_114701,IEDB_130422,IEDB_136045,IEDB_136105,IEDB_137477,IEDB_142488,IEDB_142489,IEDB_144562,IEDB_144998,IEDB_146664,IEDB_152214,IEDB_167188,IEDB_174332,IEDB_174333,IEDB_189517,IEDB_225177,IEDB_742521,IEDB_885823,IEDB_983931,SB_192,SB_86
Methods: methylation, PCR, DNA techniques, MALDI-TOF MS, radiolabeling, Southern blotting, extraction, cell growth, cloning plasmids, autoradiography, 2D-TLC
Enzymes that release or process the structure: glycosyltransferase, NAD dependent epimerase/dehydratase, glucose-1-phosphate cytidiltransferase, rhamnose epimerase, methyltransferase, polyketide synthase, fatty acyl co-A synthatase, rhamnosyltransferase, acyl transferase, fatty acyl co-A reductase, keto acyl reductase
Related record ID(s): 49318, 49320
NCBI Taxonomy refs (TaxIDs): 557599
Show glycosyltransferases
There is only one chemically distinct structure:
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Nataraj V, Pang PC, Haslam SM, Veerapen N, Minnikin DE, Dell A, Besra GS, Bhatt A
MKAN27435 is required for the biosynthesis of higher subclasses of lipooligosaccharides in Mycobacterium kansasii
PLoS One 10(3) (2015)
ID e0122804
|
LIP-(1-6)-+
|
LIP-(1-3)-+ |
| |
Lac2Me-(1-3)-b-D-QuipNMe-(1-3)-a-L-Fucp-(1-4)-{{{-b-L-Xylp-(1-4)-}}}/n=6/-a-L-Rhap2Ac3Me-(1-3)-b-D-Glcp-(1-4)-b-D-Glcp-(1-4)-a-D-Glcp-(1-1)-a-D-Glcp
|
LIP-(1-4)-+ |
Show graphically |
Mycobacterium kansasii ATCC 12478
(NCBI TaxID 557599,
species name lookup)
Taxonomic group: bacteria / Actinobacteria
(Phylum: Actinobacteria)
Associated disease: infection due to Mycobacterium kansasii [ICD11:
XN3L9 ]
NCBI PubMed ID: 25893968Publication DOI: 10.1371/journal.pone.0122804Journal NLM ID: 101285081Publisher: San Francisco, CA: Public Library of Science
Correspondence: a.bhatt
bham.ac.uk
Institutions: School of Biosciences and Institute of Microbiology and Infection, University of Birmingham, Edgbaston, UK, Department of Life Sciences, Imperial College London, London, UK
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
biosynthesis, Mycobacterium kansasii, lipooligosaccharides (LOS)
Structure type: oligomer
Location inside paper: Fig. 1, C
Compound class: lipooligosaccharide
Contained glycoepitopes: IEDB_130422,IEDB_136045,IEDB_136105,IEDB_137477,IEDB_142488,IEDB_142489,IEDB_144562,IEDB_144998,IEDB_146664,IEDB_152214,IEDB_174333,IEDB_189517,IEDB_225177,IEDB_742521,IEDB_885823,IEDB_983931,SB_192,SB_86
Methods: methylation, PCR, DNA techniques, MALDI-TOF MS, radiolabeling, Southern blotting, extraction, cell growth, cloning plasmids, autoradiography, 2D-TLC
Enzymes that release or process the structure: glycosyltransferase, polyketide synthase, fatty acyl co-A synthatase, rhamnosyltransferase, acyl transferase, fatty acyl co-A reductase, keto acyl reductase
Related record ID(s): 49318, 49319
NCBI Taxonomy refs (TaxIDs): 557599
Show glycosyltransferases
There is only one chemically distinct structure:
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