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AftD, a novel essential arabinofuranosyltransferase from mycobacteria
- Source :
- Glycobiology, Glycobiology, 2009, 19 (11), pp.1235-1247. ⟨10.1093/glycob/cwp116⟩, Glycobiology, Oxford University Press (OUP), 2009, 19 (11), pp.1235-1247. ⟨10.1093/glycob/cwp116⟩
- Publication Year :
- 2009
- Publisher :
- HAL CCSD, 2009.
-
Abstract
- International audience; Arabinogalactan (AG) and lipoarabinomannan (LAM) are the two major cell wall (lipo)polysaccharides of mycobacteria. They share arabinan chains made of linear segments of alpha-1,5-linked D-Araf residues with some alpha-1,3-branching, the biosynthesis of which offers opportunities for new chemotherapeutics. In search of the missing arabinofuranosyltransferases (AraTs) responsible for the formation of the arabinan domains of AG and LAM in Mycobacterium tuberculosis, we identified Rv0236c (AftD) as a putative membrane-associated polyprenyl-dependent glycosyltransferase. AftD is 1400 amino acid-long, making it the largest predicted glycosyltransferase of its class in the M. tuberculosis genome. Assays using cell-free extracts from recombinant Mycobacterium smegmatis and Corynebacterium glutamicum strains expressing different levels of aftD indicated that this gene encodes a functional AraT with alpha-1,3-branching activity on linear alpha-1,5-linked neoglycolipid acceptors in vitro. The disruption of aftD in M. smegmatis resulted in cell death and a decrease in its activity caused defects in cell division, reduced growth, alteration of colonial morphology, and accumulation of trehalose dimycolates in the cell envelope. Overexpression of aftD in M. smegmatis, in contrast, induced the accumulation of two arabinosylated compounds with carbohydrate backbones reminiscent of that of LAM and a degree of arabinosylation dependent on aftD expression levels. Altogether, our results thus indicate that AftD is an essential AraT involved in the synthesis of the arabinan domain of major mycobacterial cell envelope (lipo)polysaccharides.
- Subjects :
- Lipopolysaccharides
Molecular Sequence Data
Mycobacterium smegmatis
Galactans
Biochemistry
Corynebacterium glutamicum
Cell wall
03 medical and health sciences
chemistry.chemical_compound
Arabinogalactan
Glycosyltransferase
Carbohydrate Conformation
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
MESH: Carbohydrate Conformation
ComputingMilieux_MISCELLANEOUS
MESH: Mycobacterium smegmatis
030304 developmental biology
0303 health sciences
MESH: Carbohydrate Sequence
Lipoarabinomannan
MESH: Molecular Sequence Data
biology
030306 microbiology
Glycosyltransferases
MESH: Galactans
biology.organism_classification
3. Good health
Trehalose dimycolate
Carbohydrate Sequence
chemistry
MESH: Glycosyltransferases
biology.protein
Original Article
Cell envelope
MESH: Lipopolysaccharides
Subjects
Details
- Language :
- English
- ISSN :
- 09596658 and 14602423
- Database :
- OpenAIRE
- Journal :
- Glycobiology, Glycobiology, 2009, 19 (11), pp.1235-1247. ⟨10.1093/glycob/cwp116⟩, Glycobiology, Oxford University Press (OUP), 2009, 19 (11), pp.1235-1247. ⟨10.1093/glycob/cwp116⟩
- Accession number :
- edsair.doi.dedup.....cf74a7f5b47911feace459bb709a4dcb