AftD, a novel essential arabinofuranosyltransferase from mycobacteria
| Autor: | Delphi Chatterjee, Stéphanie Guadagnini, Jérôme Nigou, Patrick J. Brennan, Henrieta Škovierová, Gerald Larrouy-Maumus, Germain Puzo, Marie-Christine Prévost, Martina Belanova, Devinder Kaur, Nathalie Barilone, Mary Jackson, Brigitte Gicquel, Martine Gilleron, Jana Korduláková, Jian Zhang |
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| Přispěvatelé: | Colorado State University [Fort Collins] (CSU), Institut de pharmacologie et de biologie structurale (IPBS), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Génétique mycobactérienne - Mycobacterial genetics, Institut Pasteur [Paris] (IP), Comenius University in Bratislava, Microscopie électronique (Plate-forme), The National Institute of Allergy and Infectious Diseases/National Institutes of Health (AI064798 and AI018357), and the Slovak Research and Development Agency (APVV-0499-07)., We gratefully acknowledge A. Amin (Colorado State University) for the preparation of p[14C]Rpp, Dr. A. Dasgupta (Institut Pasteur, Paris) for his help with plasmid constructs, and Dr. P. Seeberger (Swiss Federal Institute of Technology, Zürich, Switzerland) for providing us with the synthetic Ara3, Ara4, and Man5 linear acceptor, Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Department of Microbiology, Immunology and Pathology, University of Colorado [Boulder], Institut Pasteur [Paris], Faculty of Natural Sciences |
| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
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 |
| Zdroj: | 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⟩ |
| ISSN: | 0959-6658 1460-2423 |
| DOI: | 10.1093/glycob/cwp116⟩ |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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