Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing.
Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, β1,4-galactosyltransferase I, β1,3-galactosyltransferase II, and β1,3-glucuronyltransferase. This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes.
Department of Life Science, Rikkyo University, Toshima-ku, Tokyo, Japan.|Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.
Miki
M
Yamamoto-Hino
Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.|Department of Bioinformatics, Faculty of Engineering, Soka University, Hachioji, Tokyo, Japan.|Department of Applied Biology, Insect Biomedical Research Center, Kyoto Institute of Technology, Sakyo-ku, Kyoto, Japan.
Hideki
H
Yoshida
Department of Bioinformatics, Faculty of Engineering, Soka University, Hachioji, Tokyo, Japan.
Tomomi
T
Ichimiya
Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan.
Sho
S
Sakamura
Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan.
Megumi
M
Maeda
Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan.
Yoshinobu
Y
Kimura
Department of Bioinformatics, Faculty of Engineering, Soka University, Hachioji, Tokyo, Japan.|Research Team for Geriatric Medicine (Vascular Medicine), Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo, Japan.
Norihiko
N
Sasaki
Department of Bioinformatics, Faculty of Engineering, Soka University, Hachioji, Tokyo, Japan.
Kiyoko F
KF
Aoki-Kinoshita
Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.|College of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga, Japan.
Akiko
A
Kinoshita-Toyoda
Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.|College of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga, Japan.
Hidenao
H
Toyoda
Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.|Invertebrate Genetics Laboratory, National Institute of Genetics, Mishima, Shizuoka, Japan.
Ryu
R
Ueda
Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.|Department of Bioinformatics, Faculty of Engineering, Soka University, Hachioji, Tokyo, Japan.
Shoko
S
Nishihara
Department of Life Science, Rikkyo University, Toshima-ku, Tokyo, Japan.|Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.
Satoshi
S
Goto
D000818:Animals; D004330:Drosophila; D020868:Gene Silencing; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D010641:Phenotype; D011134:Polysaccharides / Q000235:genetics; D034622:RNA Interference
D011134:Polysaccharides; D016695:Glycosyltransferases