Proteoglycan UDP-galactose:beta-xylose beta 1,4-galactosyltransferase I is essential for viability in Drosophila melanogaster.
Heparan and chondroitin sulfates play essential roles in growth factor signaling during development and share a common linkage tetrasaccharide structure, GlcAbeta1,3Galbeta1,3Galbeta1,4Xylbeta1-O-Ser. In the present study, we identified the Drosophila proteoglycan UDP-galactose:beta-xylose beta1,4-galactosyltransferase I (dbeta4GalTI), and determined its substrate specificity. The enzyme transferred a Gal to the -beta-xylose (Xyl) residue, confirming it to be the Drosophila ortholog of human proteoglycan UDP-galactose:beta-xylose beta1,4-galactosyltransferase I. Then we established UAS-dbeta4GalTI-IR fly lines containing an inverted repeat of dbeta4GalTI ligated to the upstream activating sequence (UAS) promoter, a target of GAL4, and observed the F(1) generation of the cross between the UAS-dbeta4GalTI-IR fly and the Act5C-GAL4 fly. In the F(1), double-stranded RNA of dbeta4GalTI is expressed ubiquitously under the control of a cytoplasmic actin promoter to induce the silencing of the dbeta4GalTI gene. The expression of the target gene was disrupted specifically, and the degree of interference was correlated with phenotype. The lethality among the progeny proved that beta4GalTI is essential for viability. This study is the first to use reverse genetics, RNA interference, to study the Drosophila glycosyltransferase systematically.
Takemae
Hitoshi
H
Division of Cell Biology, Soka University, Hachioji, Tokyo 192-8577, Japan.
Ueda
Ryu
R
Okubo
Reiko
R
Nakato
Hiroshi
H
Izumi
Susumu
S
Saigo
Kaoru
K
Nishihara
Shoko
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000502:physiology*; D008969:Molecular Sequence Data; D009237:N-Acetyllactosamine Synthase / Q000502:physiology*; D011509:Proteoglycans / Q000502:physiology*; D034622:RNA Interference; D014531:Uridine Diphosphate Galactose / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism*
D011509:Proteoglycans; D014531:Uridine Diphosphate Galactose; D014994:Xylose; D009237:N-Acetyllactosamine Synthase