HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 281, Issue 23, 15653-15661, June 9, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/23/15653    most recent M600045200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Alderwick, L. J. Articles by Eggeling, L. Search for Related Content PubMed PubMed Citation Articles by Alderwick, L. J. Articles by Eggeling, L. Social Bookmarking                      What's this?

Identification of a Novel Arabinofuranosyltransferase (AftA) Involved in Cell Wall Arabinan Biosynthesis in Mycobacterium tuberculosis^*

Luke J. Alderwick¹², Mathias Seidel¹, Hermann Sahm³, Gurdyal S. Besra⁴, and Lothar Eggeling³

From the School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom and Institute for Biotechnology 1, Research Centre Juelich, D-52425 Juelich, Germany

The cell wall mycolyl-arabinogalactan-peptidoglycan complex is essential in mycobacterial species, such as Mycobacterium tuberculosis, and is the target of several anti-tubercular drugs. For instance, ethambutol targets arabinogalactan biosynthesis through inhibition of the arabinofuranosyltransferases Mt-EmbA and Mt-EmbB. Following a detailed bioinformatics analysis of genes surrounding the conserved emb locus, we present the identification and characterization of a novel arabinofuranosyltransferase AftA (Rv3792). The enzyme catalyzes the addition of the first key arabinofuranosyl residue from the sugar donor -D-arabinofuranosyl-1-monophosphoryldecaprenol to the galactan domain of the cell wall, thus "priming" the galactan for further elaboration by the arabinofuranosyltransferases. Because aftA is an essential gene in M. tuberculosis, we deleted its orthologue in Corynebacterium glutamicum to produce a slow growing but viable mutant. Analysis of its cell wall revealed the complete absence of arabinose resulting in a truncated cell wall structure possessing only a galactan core with a concomitant loss of cell wall-bound mycolates. Complementation of the mutant was fully restored to the wild type phenotype by Cg-aftA. In addition, by developing an in vitro assay using recombinant Escherichia coli expressing Mt-aftA and use of cell wall galactan as an acceptor, we demonstrated the transfer of arabinose from -D-arabinofuranosyl-1-monophosphoryldecaprenol to galactan, and unlike the Mt-Emb proteins, Mt-AftA was not inhibited by ethambutol. This newly discovered glycosyltransferase represents an attractive drug target for further exploitation by chemotherapeutic intervention.

Received for publication, January 3, 2006 , and in revised form, March 31, 2006.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally to this work.

² Biotechnology and Biological Sciences Research Council Quota Student.

³ Supported by the Fonds der Chemischen Industrie.

⁴ Supported as a Lister Institute-Jenner Research Fellow and the Medical Research Council (UK). To whom correspondence should be addressed. Tel.: 121-415-8125; Fax: 121-414-5925; E-mail: g.besra{at}bham.ac.uk.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				X. Meniche, C. de Sousa-d'Auria, B. Van-der-Rest, S. Bhamidi, E. Huc, H. Huang, D. De Paepe, M. Tropis, M. McNeil, M. Daffe, et al. Partial redundancy in the synthesis of the D-arabinose incorporated in the cell wall arabinan of Corynebacterineae Microbiology, August 1, 2008; 154(8): 2315 - 2326. [Abstract] [Full Text] [PDF]

				L. Shi, R. Zhou, Z. Liu, T. L. Lowary, P. H. Seeberger, B. L. Stocker, D. C. Crick, K.-H. Khoo, and D. Chatterjee Transfer of the First Arabinofuranose Residue to Galactan Is Essential for Mycobacterium smegmatis Viability J. Bacteriol., August 1, 2008; 190(15): 5248 - 5255. [Abstract] [Full Text] [PDF]

				R. Goude, A. G. Amin, D. Chatterjee, and T. Parish The Critical Role of embC in Mycobacterium tuberculosis J. Bacteriol., June 15, 2008; 190(12): 4335 - 4341. [Abstract] [Full Text] [PDF]

				S. Bhamidi, M. S. Scherman, C. D. Rithner, J. E. Prenni, D. Chatterjee, K.-H. Khoo, and M. R. McNeil The Identification and Location of Succinyl Residues and the Characterization of the Interior Arabinan Region Allow for a Model of the Complete Primary Structure of Mycobacterium tuberculosis Mycolyl Arabinogalactan J. Biol. Chem., May 9, 2008; 283(19): 12992 - 13000. [Abstract] [Full Text] [PDF]

				D. J. Lea-Smith, K. L. Martin, J. S. Pyke, D. Tull, M. J. McConville, R. L. Coppel, and P. K. Crellin Analysis of a New Mannosyltransferase Required for the Synthesis of Phosphatidylinositol Mannosides and Lipoarbinomannan Reveals Two Lipomannan Pools in Corynebacterineae J. Biol. Chem., March 14, 2008; 283(11): 6773 - 6782. [Abstract] [Full Text] [PDF]

				E. A. Mahrous, R. B. Lee, and R. E. Lee A rapid approach to lipid profiling of mycobacteria using 2D HSQC NMR maps J. Lipid Res., February 1, 2008; 49(2): 455 - 463. [Abstract] [Full Text] [PDF]

				M. Belanova, P. Dianiskova, P. J. Brennan, G. C. Completo, N. L. Rose, T. L. Lowary, and K. Mikusova Galactosyl Transferases in Mycobacterial Cell Wall Synthesis J. Bacteriol., February 1, 2008; 190(3): 1141 - 1145. [Abstract] [Full Text] [PDF]

				D. Kaur, M. R. McNeil, K.-H. Khoo, D. Chatterjee, D. C. Crick, M. Jackson, and P. J. Brennan New Insights into the Biosynthesis of Mycobacterial Lipomannan Arising from Deletion of a Conserved Gene J. Biol. Chem., September 14, 2007; 282(37): 27133 - 27140. [Abstract] [Full Text] [PDF]

				R. V. V. Tatituri, L. J. Alderwick, A. K. Mishra, J. Nigou, M. Gilleron, K. Krumbach, P. Hitchen, A. Giordano, H. R. Morris, A. Dell, et al. Structural characterization of a partially arabinosylated lipoarabinomannan variant isolated from a Corynebacterium glutamicum ubiA mutant Microbiology, August 1, 2007; 153(8): 2621 - 2629. [Abstract] [Full Text] [PDF]

				S. Berg, D. Kaur, M. Jackson, and P. J Brennan The glycosyltransferases of Mycobacterium tuberculosis--roles in the synthesis of arabinogalactan, lipoarabinomannan, and other glycoconjugates Glycobiology, June 1, 2007; 17(6): 35R - 56R. [Abstract] [Full Text] [PDF]

				M. Seidel, L. J. Alderwick, H. L. Birch, H. Sahm, L. Eggeling, and G. S. Besra 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 J. Biol. Chem., May 18, 2007; 282(20): 14729 - 14740. [Abstract] [Full Text] [PDF]

				M. Seidel, L. J. Alderwick, H. Sahm, G. S. Besra, and L. Eggeling Topology and mutational analysis of the single Emb arabinofuranosyltransferase of Corynebacterium glutamicum as a model of Emb proteins of Mycobacterium tuberculosis Glycobiology, February 1, 2007; 17(2): 210 - 219. [Abstract] [Full Text] [PDF]

				E. Krings, K. Krumbach, B. Bathe, R. Kelle, V. F. Wendisch, H. Sahm, and L. Eggeling Characterization of myo-Inositol Utilization by Corynebacterium glutamicum: the Stimulon, Identification of Transporters, and Influence on L-Lysine Formation J. Bacteriol., December 1, 2006; 188(23): 8054 - 8061. [Abstract] [Full Text] [PDF]

				L. J. Alderwick, L. G. Dover, M. Seidel, R. Gande, H. Sahm, L. Eggeling, and G. S. Besra Arabinan-deficient mutants of Corynebacterium glutamicum and the consequent flux in decaprenylmonophosphoryl-D-arabinose metabolism Glycobiology, November 1, 2006; 16(11): 1073 - 1081. [Abstract] [Full Text] [PDF]