Approach for functional analysis of glycan using RNA interference.
The elucidation of the biological role of glycan is one of the most important issues to be resolved following the genome project. RNA interference is becoming an efficient reverse genetic tool for studying gene function in model organisms, including C.elegans and Drosophila melanogaster. Our molecular evolutionary study has shown that a prototype of glycosyltransferases, which synthesize a variety of glycan structures in the Golgi apparatus, was conserved between mammals and Drosophila. For analyses of the basic physiological functions of glycans, we established the Drosophila inducible RNAi knockdown system and applied it to one glycosyltransferase and one transporter, proteoglycan UDP-galactose: beta-xylose beta1,4galactosyltransferase I and the PAPS-transporter, respectively. If on the silencing of each gene induced ubiquitously under the control of a cytoplasmic actin promoter, the RNAi knockdown fly died, then the protein was indispensable for life. The expression of the target gene was disrupted specifically and the degree of interference was well correlated with the phenotype. The inducible RNAi knockdown fly obtained using the GAL4-UAS system will pave the way for the functional analysis of glycans.
Nishihara
Shoko
S
Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, Japan. shoko@t.soka.ac.jp
Ueda
Ryu
R
Goto
Satoshi
S
Toyoda
Hidenao
H
Ishida
Hideki
H
Nakamura
Mitsuru
M
D000199:Actins / Q000235:genetics; D000818:Animals; D001692:Biological Transport; D017173:Caenorhabditis elegans; D002241:Carbohydrates / Q000737:chemistry; D003593:Cytoplasm / Q000378:metabolism; D019992:Databases as Topic; D004331:Drosophila melanogaster; D006056:Golgi Apparatus / Q000378:metabolism; D008957:Models, Genetic; D010802:Phylogeny; D011134:Polysaccharides / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D013379:Substrate Specificity
D000199:Actins; D002241:Carbohydrates; D011134:Polysaccharides; D012333:RNA, Messenger