The nuclear envelope (NE) contains a specialized set of integral membrane proteins that maintain nuclear shape and integrity and influence chromatin organization and gene expression. Advances in proteomics techniques and studies in model organisms have identified hundreds of proteins that localize to the NE. However, the function of many of these proteins at the NE remains unclear, in part due to a lack of understanding of the interactions that these proteins participate in at the NE membrane. To assist in the characterization of NE transmembrane protein interactions we developed an arrayed library of integral and peripheral membrane proteins from the fission yeast Schizosaccharomyces pombe for high-throughput screening using the split-ubiquitin based membrane yeast two -hybrid system. We used this approach to characterize protein interactions for three conserved proteins that localize to the inner nuclear membrane: Cut11/Ndc1, Lem2 and Ima1/Samp1/Net5. Additionally, we determined how the interaction network for Cut11 is altered in canonical temperature-sensitive cut11-ts mutants. This library and screening approach is readily applicable to characterizing the interactomes of integral membrane proteins localizing to various subcellular compartments.

核膜（NE）包含一组特殊的完整膜蛋白，可以维持核的形状和完整性并影响染色质的组织和基因表达。蛋白质组学技术的发展和对模型生物的研究已经鉴定出数百种定位于NE的蛋白质。但是，许多蛋白质在NE上的功能仍不清楚，部分原因是由于对这些蛋白质在NE膜上参与的相互作用缺乏了解。为帮助表征NE跨膜蛋白相互作用，我们开发了裂变酵母粟酒裂殖酵母中完整和外围膜蛋白的阵列文库使用基于分裂泛素的膜酵母两种杂交系统进行高通量筛选。我们使用这种方法来表征定位于内部核膜的三个保守蛋白的蛋白相互作用：Cut11 / Ndc1，Lem2和Ima1 / Samp1 / Net5。此外，我们确定了如何在规范的温度敏感的cut11突变体中改变Cut11的相互作用网络。该文库和筛选方法可容易地用于表征定位于各种亚细胞区室的整合膜蛋白的相互作用组。