title: Mapping interactions between nuclear transport factors in living cells reveals pathways through the nuclear pore complex.
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.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: M
authors.initials: M
authors.lastname: Damelin
authors.forename: P A
authors.initials: PA
authors.lastname: Silver
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D002467:Cell Nucleus / Q000502:physiology; D004735:Energy Transfer; D005456:Fluorescent Dyes; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009685:Nuclear Envelope / Q000502:physiology* / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000502:physiology* / Q000648:ultrastructure*
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria