title: A size-exclusion permeability barrier and nucleoporins characterize a ciliary pore complex that regulates transport into cilia.
abstract: The cilium is a microtubule-based organelle that contains a unique complement of proteins for cell motility and signalling functions. Entry into the ciliary compartment is proposed to be regulated at the base of the cilium. Recent work demonstrated that components of the nuclear import machinery, including the Ran GTPase and importins, regulate ciliary entry. We hypothesized that the ciliary base contains a ciliary pore complex whose molecular nature and selective mechanism are similar to those of the nuclear pore complex. By microinjecting fluorescently labelled dextrans and recombinant proteins of various sizes, we characterize a size-dependent diffusion barrier for the entry of cytoplasmic molecules into primary cilia in mammalian cells. We demonstrate that nucleoporins localize to the base of primary and motile cilia and that microinjection of nucleoporin-function-blocking reagents blocks the ciliary entry of kinesin-2 KIF17 motors. Together, this work demonstrates that the physical and molecular nature of the ciliary pore complex is similar to that of the nuclear pore complex, and further extends functional parallels between nuclear and ciliary import.
authors.affiliation: Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
authors.forename: Hooi Lynn
authors.initials: HL
authors.lastname: Kee
authors.forename: John F
authors.initials: JF
authors.lastname: Dishinger
authors.forename: T Lynne
authors.initials: TL
authors.lastname: Blasius
authors.forename: Chia-Jen
authors.initials: CJ
authors.lastname: Liu
authors.forename: Ben
authors.initials: B
authors.lastname: Margolis
authors.forename: Kristen J
authors.initials: KJ
authors.lastname: Verhey
mesh_terms: D001692:Biological Transport; D002478:Cells, Cultured; D002923:Cilia / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D003911:Dextrans; D006801:Humans; D008970:Molecular Weight; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism*; D010539:Permeability; D021381:Protein Transport
chemical_list: D003911:Dextrans; D028861:Nuclear Pore Complex Proteins