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

J. Biol. Chem., Vol. 282, Issue 20, 14729-14740, May 18, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/20/14729    most recent M700271200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Seidel, M. Articles by Besra, G. S. Search for Related Content PubMed PubMed Citation Articles by Seidel, M. Articles by Besra, G. S. Social Bookmarking                      What's this?

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^*

Mathias Seidel¹, Luke J. Alderwick¹², Helen L. Birch, Hermann Sahm, Lothar Eggeling, and Gurdyal S. Besra³

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

Arabinofuranosyltransferase enzymes, such as EmbA, EmbB, and AftA, play pivotal roles in the biosynthesis of arabinogalactan, and the anti-tuberculosis agent ethambutol (EMB) targets arabinogalactan biosynthesis through inhibition of Mt-EmbA and Mt-EmbB. Herein, we describe the identification and characterization of a novel arabinofuranosyltransferase, now termed AftB (Rv3805c), which is essential in Mycobacterium tuberculosis. Deletion of its orthologue NCgl2780 in the closely related species Corynebacterium glutamicum resulted in a viable mutant. Analysis of the cell wall-associated lipids from the deletion mutant revealed a decreased abundance of cell wall-bound mycolic acids, consistent with a partial loss of mycolylation sites. Subsequent glycosyl linkage analysis of arabinogalactan also revealed the complete absence of terminal (1 2)-linked arabinofuranosyl residues. The deletion mutant biochemical phenotype was fully complemented by either Mt-AftB or Cg-AftB, but not with muteins of Mt-AftB, where the two adjacent aspartic acid residues, which have been suggested to be involved in glycosyltransferase activity, were replaced by alanine. In addition, the use of C. glutamicum and C. glutamicumaftB in an in vitro assay utilizing the sugar donor -D-arabinofuranosyl-1-monophosphoryl-decaprenol together with the neoglycolipid acceptor -D-Araf-(1 5)--D-Araf-O-C₈ as a substrate confirmed AftB as a terminal (1 2) arabinofuranosyltransferase, which was also insensitive to EMB. Altogether, these studies have shed further light on the complexities of Corynebacterianeae cell wall biosynthesis, and Mt-AftB represents a potential new drug target.

Received for publication, January 10, 2007 , and in revised form, February 16, 2007.

^* This work was supported by the Medical Research Council (UK), the Wellcome Trust, and by the Fonds der Chemischen Industrie for support (to H. S.). 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 These authors contributed equally to this work.

² A Biotechnology and Biological Sciences Research Council Quota student.

³ 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:

				A. K. Mishra, S. Batt, K. Krumbach, L. Eggeling, and G. S. Besra Characterization of the Corynebacterium glutamicum {Delta}pimB' {Delta}mgtA Double Deletion Mutant and the Role of Mycobacterium tuberculosis Orthologues Rv2188c and Rv0557 in Glycolipid Biosynthesis J. Bacteriol., July 1, 2009; 191(13): 4465 - 4472. [Abstract] [Full Text] [PDF]

				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]

				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]

				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]