JOUBERIN AND NEPHROCYSTIN IN JOUBERT SYNDROME

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 Joubert综合征（JS）是一种以小脑蚓部发育不全、眼球运动异常、共济失调和智力低下为特征的脑发育障碍。JS通常涉及与几种纤毛相关疾病显著重叠的CNS外表型，暗示JS中的纤毛功能障碍。CEP 290是JS中突变的几个基因之一。CEP 290中的无义突变或移码突变是造成大部分眼肾型JS的原因，而CEP 290中产生强剪接供体位点的内含子突变是Leber先天性黑蒙（一种严重的视网膜营养不良）的单一最常见的确定原因。CEP 290是具有原肌球蛋白同源结构域的大卷曲螺旋蛋白。已知CEP 290定位于纤毛肾细胞的中心体和基体以及视网膜光感受器的连接纤毛。此外，CEP 290从hTERT-RPE细胞的耗竭被证明干扰血清饥饿诱导的纤毛发生。然而，CEP 290在基体中的功能的分子机制在很大程度上仍然未知。在本研究中，我们使用串联亲和纯化从hTERT-RPE细胞中分离CEP 290蛋白复合物。通过质谱鉴定复合物中的蛋白质将为理解CEP 290的功能和JS的病理机制提供重要信息。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Joubert syndrome (JS) is a developmental brain disorder characterized by cerebellar vermis hypoplasia, abnormal eye movement, ataxia and mental retardation. JS often involves extra-CNS phenotypes which significantly overlap with several cilia-related disorders, implicating ciliary dysfunction in JS. CEP290 is one of several genes mutated in JS. Nonsense or frame-shift mutations in CEP290 are responsible for a majority of the oculorenal form of JS, whereas an intronic mutation in CEP290 that creates a strong splice-donor site is the single most common identified cause of Leber congenital amaurosis, a severe retinal dystrophy. CEP290 is a large coiled-coil protein with tropomyosin homology domains. CEP290 is known to localize to the centrosome and basal body of ciliated kidney cells and to the connecting cilium of retinal photoreceptors. Moreover, depletion of CEP290 from hTERT-RPE cells was shown to interfere with serum starvation-induced ciliogenesis. However, molecular mechanism of CEP290 function at the basal body remains largely unknown. In this study, we use tandem affinity purification to isolate CEP290 protein complex from hTERT-RPE cells. Identification of proteins in the complex by mass spectrometry will provide vital information for understanding both CEP290 function and the pathological mechanism of JS.