EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorhabditis elegans

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• 作者：Morio ; Hidenori ; Honda ; Yoko ; Toyoda ; Hidenao ; Nakajima ; Mitsunari ; Kurosawa ; Hisashi ; Shirasawa ; Takuji
• 关键词：Caenorhabditis elegans ; rib-2 ; Heparan sulfate ; Egg-laying defect ; Developmental defect
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2003
• 出版时间：February 7, 2003
• 年：2003
• 卷：301
• 期：2
• 页码：317-323
• 全文大小：253 K

文摘

EXT gene family members including EXT1, EXT2, and EXTL2 are glycosyltransferases required for heparan sulfate biosynthesis. To examine the biological functions of rib-2, a member of the Caenorhabditis elegans EXT gene family, we generated a mutant worm lacking the rib-2 gene using the UV-TMP method followed by sib-selection. Inactivation of rib-2 alleles induced developmental abnormalities in F2 and F3 homozygous worms, while F1 heterozygotes showed a normal morphology. The F2 homozygous progeny generated from the F1 heterozygous hermaphrodites somehow developed to adult stage but exhibited abnormal characteristics such as developmental delay and egg-laying defects. The F3 homozygous progeny from the F2 homozygous hermaphrodites showed early developmental defects and most of the F3 worms stopped developing during the gastrulation stage. Whole-mount staining analysis for heparan sulfate using Toluidine blue (pH 2.5) revealed a defect of heparan sulfate biosynthesis in the F2 homozygotes. The analysis using fluorometric post-column high-performance liquid chromatography also uncovered reduced production of heparan sulfate in the rib-2 mutant. These results indicate that rib-2 is essential for embryonic development and heparan sulfate biosynthesis in C. elegans.