Human Molecular Genetics Advance Access originally published online on January 20, 2005
Human Molecular Genetics 2005 14(5):657-665; doi:10.1093/hmg/ddi062
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Human Molecular Genetics, Vol. 14, No. 5 © Oxford University Press 2005; all rights reserved
Localization and functional analysis of the LARGE family of glycosyltransferases: significance for muscular dystrophy
Dubowitz Neuromuscular Centre, Department of Paediatrics, Hammersmith Campus, Imperial College, Du Cane Road, London W12 0NN, UK
* To whom correspondence should be addressed. Tel: +44 2083833148; Fax: +44 2087462187; Email: f.muntoni{at}imperial.ac.uk
Received October 3, 2004; Revised December 12, 2004; Accepted January 12, 2005
The dystroglycanopathies are a novel group of human muscular dystrophies due to mutations in known or putative glycosyltransferase enzymes. They share the common pathological feature of a hypoglycosylated form of 
-dystroglycan, diminishing its ability to bind extracellular matrix ligands. The LARGE glycosyltransferase is mutated in both the myodystrophy mouse and congenital muscular dystrophy type 1D (MDC1D). We have transfected various cell lines with a variety of LARGE expression constructs in order to characterize their subcellular localization and effect on -dystroglycan glycosylation. Wild-type LARGE co-localized with the Golgi marker GM130 and stimulated the production of highly glycosylated -dystroglycan (hyperglycosylation). MDC1D mutants had no effect on -dystroglycan glycosylation and failed to localize correctly, confirming their pathogenicity. The two predicted catalytic domains of LARGE contain three conserved DxD motifs. Systematically mutating each of these motifs to NNN resulted in the mislocalization of one construct, while all failed to have any effect on -dystroglycan glycosylation. A construct lacking the transmembrane domain also failed to localize at the Golgi apparatus. These results indicate that LARGE needs to both physically interact with -dystroglycan and function as a glycosyltransferase in order to stimulate -dystroglycan hyperglycosylation. We have also cloned and overexpressed a homologue of LARGE, glycosyltransferase-like 1B (GYLTL1B). Like LARGE it localized to the Golgi apparatus and stimulated -dystroglycan hyperglycosylation. These results suggest that GYLTL1B may be a candidate gene for muscular dystrophy and that its overexpression could compensate for the deficiency of both LARGE and other glycosyltransferases.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
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