JOUBERIN AND NEPHROCYSTIN IN JOUBERT SYNDROME

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 Joubert综合征(JS)是一种以小脑蠕虫发育不全、眼球运动异常、共济失调和智力低下为特征的发育性脑疾病。JS常涉及CNS外表型，与几种纤毛相关疾病显著重叠，提示JS存在纤毛功能障碍。CEP290是JS中突变的几个基因之一。CEP290的无义或移码突变是大多数眼肾形JS的原因，而CEP290的内含子突变产生了一个强大的剪接供体部位，是Leber先天性黑色素的最常见原因，Leber先天性黑色素是一种严重的视网膜营养不良。CEP290是一种具有原肌球蛋白同源结构域的大卷曲蛋白。已知CEP290定位于有纤毛的肾细胞的中心体和基底，以及视网膜光感受器的连接纤毛。此外，hTERT-RPE细胞中CEP290的缺失被证明干扰了血清饥饿诱导的纤毛发生。然而，CEP290在基底体发挥作用的分子机制仍很不清楚。在本研究中，我们采用串联亲和纯化的方法从hTERT-RPE细胞中分离出CEP290蛋白复合体。用质谱仪鉴定该复合体中的蛋白质将为了解CEP290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.