Analysis of the Nup214-CRM1 interaction in nuclear protein export

核孔蛋白Nup 214是核孔复合物的一个组成部分，最初被鉴定为与其他蛋白质融合的潜在癌基因。最近，发现Nup 214在乳腺癌肿瘤细胞中突变，其突变率证明其被归类为“癌症基因”是合理的。已知Nup 214在核质转运中起作用，然而，对于由于突变的Nup 214而促进肿瘤发生的可能机制一无所知。一个有吸引力的假设，这是由我们和其他人发表的工作支持，是Nup 214所需的选择蛋白质的核输出与生长控制的联系。一个这样的候选者是转录因子NFAT（活化T细胞的核因子），其CRM 1依赖性输出需要Nup 214。在这里，我们建议调查Nup 214的功能，在肿瘤发生的可能参与，其中一个子集的蛋白质的核出口可能会受到抑制。对于我们的分析，我们将使用我们建立的方案通过RNA干扰来耗尽Nup 214。此外，我们将利用来自模式生物C的转运因子。elegans来分析CRM 1、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Nup 214中的突变/缺失及其在乳腺癌细胞中对NFAT和其他货物蛋白的核转运的可能后果。总之，我们希望提高我们对Nup 214在生理和病理条件下的功能的理解。-

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. -