Functional Analysis of Diastrophic Dysplasia Sulfate Transporter
Functional Analysis of Diastrophic Dysplasia Sulfate Transporter
复制标题
发育不良硫酸盐转运蛋白的功能分析
DOI：
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发表时间：
1998
影响因子：
4.8
通讯作者：
Y. Hiraki
中科院分区：
文献类型：
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作者：
Hideshi Satoh;Masakazu Susaki;C. Shukunami;K. Iyama;T. Negoro;Y. Hiraki
骨畸形性发育不良硫酸盐转运蛋白（DTDST）基因突变构成了一个家族的reception-inherited骨软骨发育不良，包括软骨发育不全型1B，骨发育不全型II，和骨畸形性发育不良。然而，基因产物的功能特性尚未阐明。我们从大鼠UMR-106成骨细胞中克隆了大鼠DTDST cDNA。北方印迹分析表明，软骨和小肠是DTDST mRNA的主要表达部位。基因组序列分析表明，大鼠DTDST基因由至少5个外显子组成。两种不同的转录本在软骨细胞中表达，这是由于第三个外显子的交替利用，对应于cDNA的5′-非翻译区的内部部分。将大鼠和人DTDST cRNA注射到非洲爪蟾卵母细胞中诱导Na+非依赖性硫酸盐转运。表达的DTDST的转运活性被细胞外的氯和碳酸氢盐显着抑制。相比之下，小管Na+-非依赖性硫酸盐转运蛋白Sat-1需要在注射cRNA的卵母细胞中存在胞外氯。在生长板软骨细胞中的硫酸盐运输的活性分布进行了研究，在细胞外存在的各种阴离子，并发现基本上相同的卵母细胞中表达的DTDST。因此，软骨细胞的硫酸盐转运主要依赖于DTDST系统。最后，我们表明，硫酸化不足的蛋白多糖的氯酸盐治疗的软骨细胞显着受损的细胞成纤维细胞生长因子的生长反应，这表明DTDST在endochondrium骨形成的作用。
Mutations in the diastrophic dysplasia sulfate transporter (DTDST) gene constitute a family of recessively inherited osteochondrodysplasias including achondrogenesis type 1B, atelosteogenesis type II, and diastrophic dysplasia. However, the functional properties of the gene product have yet to be elucidated. We cloned rat DTDST cDNA from rat UMR-106 osteoblastic cells. Northern blot analysis suggested that cartilage and intestine were the major expression sites for DTDST mRNA. Analysis of the genomic sequence revealed that the rat DTDST gene was composed of at least five exons. Two distinct transcripts were expressed in chondrocytes due to alternative utilization of the third exon, corresponding to an internal portion of the 5′-untranslated region of the cDNA. Injection of rat and human DTDST cRNA into Xenopus laevis oocytes induced Na+-independent sulfate transport. Transport activity of the expressed DTDST was markedly inhibited by extracellular chloride and bicarbonate. In contrast, canalicular Na+-independent sulfate transporter Sat-1 required the presence of extracellular chloride in the cRNA-injected oocytes. The activity profile of sulfate transport in growth plate chondrocytes was studied in the extracellular presence of various anions and found substantially identical to DTDST expressed in oocytes. Thus, sulfate transport of chondrocytes is dominantly dependent on the DTDST system. Finally, we demonstrate that undersulfation of proteoglycans by the chlorate treatment of chondrocytes significantly impaired growth response of the cells to fibroblast growth factor, suggesting a role for DTDST in endochondral bone formation.
影响因子：
9.8
作者：
J. Hästbacka;A. Superti-Furga;W. Wilcox;D. Rimoin;D. Cohn;E. Lander
通讯作者：
J. Hästbacka;A. Superti-Furga;W. Wilcox;D. Rimoin;D. Cohn;E. Lander
大鼠骨肉瘤的成骨细胞样细胞的腺苷酸环化酶受到降钙素和甲状旁腺激素的刺激。
影响因子：
4.8
作者：
Gutierrez,GE;Mundy,GR;Katz,MS
通讯作者：
Katz,MS