Studies on the biosynthetic mechanism and regulation of the expression of sulfated glycosaminoglycans

硫酸化糖胺聚糖，包括肝素/硫酸乙酰肝素和软骨素/硫酸皮肤素，已经涉及脊椎动物和无脊椎动物的许多病理生理学现象。在这项研究中，我们发现硫酸化糖胺聚糖的新功能如下：1）我们确定了脊椎骨骺发育不良阿曼型的致病基因为CHST 3，编码软骨素6-O-磺基转移酶-1（C6 ST-1）。C6 ST-1通过将硫酸根转移到软骨素的GalNAc的C-6位来催化硫酸软骨素合成的修饰步骤。研究结果表明CHST 3突变导致硫酸软骨素链硫酸化的特异性但普遍性缺陷，导致主要累及脊柱的软骨发育不良。2）我们克隆了秀丽隐杆线虫软骨素聚合因子（cChPF）。在cChPF被RNAi耗尽后观察到的蠕虫表型包括胞质分裂的逆转，与秀丽隐杆线虫软骨素合酶（cChSy）-RNAi phen非常相似。 ...更多信息 类型。因此，cChPF除了cChSy是必不可少的软骨素的生物合成和胚胎细胞分裂在秀丽隐杆线虫。3）我们发现，硫酸软骨素的特征在于E-二糖单位是一种有效的抑制剂的单纯疱疹病毒的感染性，并提供病毒的结合位点的gro 2C细胞。了解硫酸软骨素-E的抗病毒特性的决定因素将有助于开发人类单纯疱疹病毒感染的抑制剂。4）硫酸乙酰肝素（HS）链的形成由EXT（遗传性多外生骨素基因）家族成员编码的糖基转移酶催化。在果蝇中，已经鉴定了三个EXT家族基因，分别命名为tout-velu（tttv）、tout-velu的姐妹基因（sotv）和tout-velu的兄弟基因（botv），它们分别编码人EXT 1、EXT 2和EXTL 3的同源物。在这里，我们证明了果蝇中的所有三个EXT成员，TTV，SOTV和BOTV，都是果蝇中全长HS生物合成所必需的。5）所有生物体中的所有硫酸化反应都需要活化的硫酸盐，PAPS作为通用供体。我们克隆了秀丽隐杆线虫PAPS合成酶基因pps-1。干扰的PPS-1基因引起多效性的发育缺陷，肌肉图案和上皮细胞形状的变化，减少糖胺聚糖硫酸化，这表明硫酸化是必不可少的完整性表皮秀丽隐杆线虫。少

Sulfated glycosaminoglycans including heparin/heparan sulfate and chondroitin/dermatan sulfate have been implicated in numerous pathophysiological phenomena of vertebrates and invertebrates. In this study, we found novel functions of sulfated glycosaminoglycans as follows.1)We identified the causal gene for the spondylepiphyseal dysplasia Omani type as CHST3 that encodes chondroitin 6-O-sulfotransferase-1 (C6ST-1). C6ST-1 catalyzes the modifying step of chondroitin sulfate synthesis by transferring sulfate to the C-6 position of the GalNAc of chondroitin. The findings indicate that the mutation in CHST3 causes a specific but generalized defect of chondroitin sulfate chain sulfation resulting in chondrodysplasia with major involvement of the spine.2)We cloned C.elegans chondroitin polymerizing factor (cChPF). The worm phenotypes including the reversion of cytokinesis, observed after the depletion of cChPF by RNAi, were very similar to the C.elegans chondroitin synthase (cChSy)-RNAi phen … More otypes. Thus, cChPF in addition to cChSy is indispensable for the biosynthesis of chondroitin and embryonic cell division in C.elegans.3)We found that chondroitin sulfate characterized by the E-disaccharide unit was a potent inhibitor of herpes simplex virus infectivity and provided the virus binding sites on gro2C cells. Knowledge of the determinants of antiviral properties of chondroitin sulfate-E will help in the development of inhibitors of herpes simplex virus infections in humans.4)The formation of heparan sulfate (HS) chains is catalyzed by glycosyltransferases encoded by EXT (hereditary multiple exostosin gene) family members. In Drosophila, three EXT family genes named tout-velu (ttv), sister of tout-velu (sotv) and brother of tout-velu (botv), which encode homologues of human EXT1, EXT2 and EXTL3, respectively, have been identified. Here, we demonstrated that all three EXT members in Drosophila, TTV, SOTV and BOTV, are required for the biosynthesis of full-length HS in Drosophila.5)All sulfation reactions in all organisms require activated sulfate, PAPS as a universal donor. We cloned a C.elegans PAPS-synthase gene pps-1. Disruption of the pps-1 gene by RNAi caused pleiotropic developmental defects in muscle patterning and epithelial cell shape changes with decrease in glycosaminoglycan sulfation, suggesting that sulfation is essential for integrity of epidermis in C.elegans. Less