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J. Biol. Chem., Vol. 281, Issue 18, 12776-12785, May 5, 2006

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Null Mutations in Drosophila N-Acetylglucosaminyltransferase I Produce Defects in Locomotion and a Reduced Life Span^*

Mohan Sarkar, Peter A. Leventis^¶, Cristina I. Silvescu^||, Vernon N. Reinhold^||, Harry Schachter^**1, and Gabrielle L. Boulianne^¶2

From the Program in Structural Biology and Biochemistry and the Program in Developmental Biology, The Hospital for Sick Ontario M5G 1X8, Canada, the ^¶Department of Zoology, University of Toronto, Toronto, Ontario M5S 3G5, Canada, the ^||Department of Chemistry, the University of New Hampshire, Durham, New Hampshire 03824, and the ^**Department of Biochemistry and the Department of Molecular and Medical Genetics, the University of Toronto, Toronto, Ontario M5S 1A8, Canada

UDP-GlcNAc:3-D-mannoside 1,2-N-acetylglucosaminyltransferase I (encoded by Mgat1) controls the synthesis of hybrid, complex, and paucimannose N-glycans. Mice make hybrid and complex N-glycans but little or no paucimannose N-glycans. In contrast, Drosophila melanogaster and Caenorhabditis elegans make paucimannose N-glycans but little or no hybrid or complex N-glycans. To determine the functional requirement for 1,2-N-acetylglucosaminyltransferase I in Drosophila, we generated null mutations by imprecise excision of a nearby transposable element. Extracts from Mgat1¹/Mgat1¹ null mutants showed no 1,2-N-acetylglucosaminyltransferase I enzyme activity. Moreover, mass spectrometric analysis of these extracts showed dramatic changes in N-glycans compatible with lack of 1,2-N-acetylglucosaminyltransferase I activity. Interestingly, Mgat1¹/Mgat1¹ null mutants are viable but exhibit pronounced defects in adult locomotory activity when compared with Mgat1¹/CyO-GFP heterozygotes or wild type flies. In addition, in null mutants males are sterile and have a severely reduced mean and maximum life span. Microscopic examination of mutant adult fly brains showed the presence of fused lobes. The removal of both maternal and zygotic Mgat1 also gave rise to embryos that no longer express the horseradish peroxidase antigen within the central nervous system. Taken together, the data indicate that 1,2-N-acetylglucosaminyltransferase I-dependent N-glycans are required for locomotory activity, life span, and brain development in Drosophila.

Received for publication, November 29, 2005 , and in revised form, February 14, 2006.

^* This work was supported by funds from the Canadian Institutes of Health Research (to H. S. and G. L. B.), Natural Science and Engineering Research Council funds (to G. L. B.), and a Canadian Institutes of Health Research doctoral research award (to P. A. L.). 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.

² Recipient of a Canada Research Chair in Molecular and Developmental Neurobiology.

¹ To whom correspondence should be addressed: Program in Structural Biology and Biochemistry, The Hospital for Sick Children, 555 University Ave., Toronto, Ontario M5G 1X8, Canada. Tel.: 416-813-5915; Fax: 416-813-5022; E-mail: harry{at}sickkids.ca.

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