title: Selectivity mechanism of the nuclear pore complex characterized by single cargo tracking.
abstract: The nuclear pore complex (NPC) mediates all exchange between the cytoplasm and the nucleus. Small molecules can passively diffuse through the NPC, whereas larger cargos require transport receptors to translocate. How the NPC facilitates the translocation of transport receptor/cargo complexes remains unclear. To investigate this process, we tracked single protein-functionalized quantum dot cargos as they moved through human NPCs. Here we show that import proceeds by successive substeps comprising cargo capture, filtering and translocation, and release into the nucleus. Most quantum dots are rejected at one of these steps and return to the cytoplasm, including very large cargos that abort at a size-selective barrier. Cargo movement in the central channel is subdiffusive and cargos that can bind more transport receptors diffuse more freely. Without Ran GTPase, a critical regulator of transport directionality, cargos still explore the entire NPC, but have a markedly reduced probability of exit into the nucleus, suggesting that NPC entry and exit steps are not equivalent and that the pore is functionally asymmetric to importing cargos. The overall selectivity of the NPC seems to arise from the cumulative action of multiple reversible substeps and a final irreversible exit step.
authors.affiliation: Department of Physics, University of California, Berkeley, California 94720, USA.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Lowe
authors.forename: Jake J
authors.initials: JJ
authors.lastname: Siegel
authors.forename: Petr
authors.initials: P
authors.lastname: Kalab
authors.forename: Merek
authors.initials: M
authors.lastname: Siu
authors.forename: Karsten
authors.initials: K
authors.lastname: Weis
authors.forename: Jan T
authors.initials: JT
authors.lastname: Liphardt
mesh_terms: D021581:Active Transport, Cell Nucleus; D003593:Cytoplasm / Q000378:metabolism; D004058:Diffusion; D006367:HeLa Cells; D006801:Humans; D008970:Molecular Weight; D009068:Movement; D022022:Nuclear Pore / Q000378:metabolism*; D029543:Nucleocytoplasmic Transport Proteins / Q000378:metabolism*; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D045663:Quantum Dots; D013379:Substrate Specificity; D020931:ran GTP-Binding Protein / Q000378:metabolism
chemical_list: D029543:Nucleocytoplasmic Transport Proteins; D011506:Proteins; D020931:ran GTP-Binding Protein
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