Mapping interactions between nuclear transport factors in living cells reveals pathways through the nuclear pore complex

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Abstract

The interactions between transport receptors and proteins of the nuclear pore complex (NPC) are fundamental to understanding nucleocytoplasmic transport. In order to delineate the path that a particular transport receptor takes through the NPC, we have employed fluorescence resonance energy transfer (FRET) between enhanced cyan and yellow fluorescent proteins (ECFP, EYFP) in living cells. A panel of yeast strains expressing functional receptor-ECFP and nucleoporin-EYFP fusions has been analyzed with a FRET assay. With this approach, we define points of contact in the NPC for the related importin Pse1/Kap121 and exportin Msn5. These data demonstrate the utility of FRET in mapping dynamic protein interactions in a genetic system. Furthermore, the data indicate that an importin and exportin have overlapping pathways through the NPC.

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Damelin, M., & Silver, P. A. (2000). Mapping interactions between nuclear transport factors in living cells reveals pathways through the nuclear pore complex. Molecular Cell, 5(1), 133–140. https://doi.org/10.1016/S1097-2765(00)80409-8

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