Mapping the native organization of the yeast nuclear pore complex using nuclear radial intensity measurements

• DOI: 10.1073/pnas.1903764116
• Publication Date: 2019-07-02T00:25:48Z
• Authors: Pascal Vallotton; Sasikumar Rajoo; Matthias Wojtynek; Evgeny Onischenko; Annemarie Kralt; Carina Patrizia D...; Karsten Weis
• Keywords: Nuclear Pore Complex Proteins; 0301 basic medicine; 03 medical and health sciences; Saccharomyces cerevisiae Proteins; Intravital Microscopy; Microscopy, Fluorescence; Nuclear pore complex; Superresolution; Nucleoporins; Quantitative; Fluorescence microscopy; FG repeats; Active Transport, Cell Nucleus; Nuclear Pore; Saccharomyces cerevisiae; Chlamydomonas reinhardtii

SignificanceThe nuclear pore complex (NPC) is a huge protein assembly that selectively transports macromolecules across the nuclear envelope (NE) of eukaryotes. Determining the precise organization of that machinery has been a long-standing goal of structural biology. Here, we introduce a methodology called NuRIM that can map the average position of NE proteins in vivo based on the analysis of intensity patterns in fluorescence micrographs. This generally applicable technique delivers a precise positional map of the native yeast NPC and associated factors. Further, it allows us to investigate the structural consequences of NPC compositional perturbations, and to orient specific protein segments that play an essential role in NPC transport and assembly.