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Kamimura, K, Maeda, N and Nakato, H (2011) In vivo manipulation of heparan sulfate structure and its effect on Drosophila development. Glycobiology 21:607-18
Heparan sulfate proteoglycans (HSPGs) participate in a wide range of biological processes through interactions with a number of ligand proteins. The nature of these interactions largely depends on the heparan sulfate (HS) moiety of HSPGs, which undergoes a series of modifications by various HS-modifying enzymes (HSMEs). Although the effects of alterations in a single HSME on physiological processes have started to be studied, it remains elusive how a combination of these molecules control the structure and function of HS. Here we systematically manipulated the HS structures and analyzed their effect on morphogenesis and signaling, using the genetically tractable model organism, Drosophila. We generated transgenic fly strains overexpressing HSMEs alone or in combination. Unsaturated disaccharide analyses of HS showed that expression of various HSMEs generates distinct HS structures, and the enzymatic activities of HSMEs are influenced by coexpression of other HSMEs. Furthermore, these transgenic HSME animals showed a different extent of lethality, and a subset of HSMEs caused specific morphological defects due to defective activities of Wnt and bone morphogenetic protein signaling. There is no obvious relationship between HS unsaturated disaccharide composition and developmental defects in HSME animals, suggesting that other structural factors, such as domain organization or sulfation sequence, might regulate the function of HS.
Animals; Animals, Genetically Modified; Carbohydrate Epimerases/biosynthesis; Carbohydrate Epimerases/genetics; Cell Proliferation; Drosophila/embryology; Drosophila/genetics; Drosophila/metabolism; Drosophila Proteins/biosynthesis; Drosophila Proteins/genetics; Drosophila Proteins/metabolism; Heparan Sulfate Proteoglycans/chemistry; Heparan Sulfate Proteoglycans/metabolism; Hybridization, Genetic; Phenotype; Recombinant Proteins/biosynthesis; Recombinant Proteins/genetics; Signal Transduction/genetics; Sulfotransferases/biosynthesis; Sulfotransferases/genetics; Transgenes; Wing/abnormalities; Wnt1 Protein/metabolism