RT Journal Article SR Electronic T1 High-throughput identification of nuclear envelope protein interactions in Schizosaccharomyces pombe using an arrayed membrane yeast-two hybrid library JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.07.29.227819 DO 10.1101/2020.07.29.227819 A1 Joseph M. Varberg A1 Jennifer M. Gardner A1 Scott McCroskey A1 Snehabala Saravanan A1 William D. Bradford A1 Sue L. Jaspersen YR 2020 UL http://biorxiv.org/content/early/2020/07/30/2020.07.29.227819.abstract AB 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 in the fission yeast Schizosaccharomyces pombe for high-throughput screening using the split-ubiquitin based membrane yeast two hybrid sys-tem. We used this approach to characterize protein interactions for three conserved proteins that localize to the inner nu-clear membrane: Cut11/Ndc1, Lem2, and Ima1/Samp1/NET5. Additionally, we determined how the interaction network for Cut11 is altered in canonical temperature-sensitive cut11 mutants. This library and screening approach is readily applicable to characterizing the interactomes of integral membrane proteins localizing to various subcellular compartments.Competing Interest StatementThe authors have declared no competing interest.