Analysis of the Nup214-CRM1 interaction in nuclear protein export

核孔蛋白Nup214是核孔复合物的一个组成部分，最初被确定为与其他蛋白质融合的潜在致癌基因。最近，Nup214被发现在乳腺癌肿瘤细胞中发生突变，其突变速率证明了其作为“癌症基因”的分类是合理的。已知Nup214在核胞质转运中发挥作用，然而，关于Nup214突变促进肿瘤发生的可能机制尚不清楚。我们和其他人发表的研究支持了一个有吸引力的假设，即Nup214是与生长控制有关的选定蛋白质的核输出所必需的。其中一个候选因子是转录因子NFAT（活化T细胞的核因子），其依赖于crm1的输出需要Nup214。在这里，我们建议研究Nup214的功能，因为它可能参与肿瘤发生，其中一个蛋白质子集的核输出可能被抑制。在我们的分析中，我们将使用我们建立的协议通过rna干扰来消耗Nup214。此外，我们将利用模式生物秀丽隐杆线虫的运输因子来分析CRM1、核孔蛋白和出口货物之间相互作用的要求。最后，我们将分析Nup214的突变/缺失及其对乳腺癌细胞中NFAT和其他货物蛋白核转运的可能影响。总之，我们希望加深我们对Nup214在生理和病理条件下功能的理解。-

The nucleoporin Nup214, a component of the nuclear pore complex, was initially identified as a potential oncogene in a fusion with other proteins. Very recently, Nup214 was found to be mutated in breast cancer tumor cells at a rate that justified its classification as a “cancer gene”. Nup214 is known to play a role in nucleocytoplasmic transport, nothing is known, however, about possible mechanisms that would promote tumorigenesis as a result of mutated Nup214. An attractive hypothesis, which is supported by published work from us and others, is that Nup214 is required for nuclear export of selected proteins with links to growth control. One such candidate is the transcription factor NFAT (nuclear factor of activated T cells), whose CRM1-dependent export requires Nup214. Here we propose to investigate the function of Nup214 in light of a possible involvement in tumorigenesis, where nuclear export of a subset of proteins might be inhibited. For our analysis, we will use our established protocols to deplete Nup214 by RNA-interference. Furthermore, we will take advantage of transport factors from the model organism C. elegans to analyze the requirements for the interaction between CRM1, the nucleoporin and an export cargo. Finally, we will analyze mutations/deletions in Nup214 and their possible consequences for nuclear transport of NFAT and other cargo proteins in the context of breast cancer cells. Together, we hope to enhance our understanding of the function of Nup214 under physiological as well as pathological conditions. -