Exo70-mediated recruitment of nucleoporin Nup62 at the leading edge of migrating cells is required for cell migration.
Nucleoporin Nup62 localizes at the central channel of the nuclear pore complex and is essential for nucleocytoplasmic transport. Through its FG-repeat domain, Nup62 regulates nuclear pore permeability and binds nuclear transport receptors. Here, we report that Nup62 interacts directly with Exo70 and colocalizes with Exo70 at the leading edge of migrating cells. Nup62 binds the N-terminal domain of Exo70 through its coiled-coil domain but not through its FG-repeat domain. Selective inhibition of leading edge Nup62 using RNA interference significantly reduces cell migration. Furthermore, Exo70 recruits Nup62 at the plasma membrane and at filopodia. Removal of the Exo70-binding domain of Nup62 prevents leading edge localization of Nup62. Analogous to Exo70, Nup62 cycles between the plasma membrane and the perinuclear recycling compartment. Altogether, we propose that Nup62 not solely regulates access to the cell nucleus, but additionally functions in conjunction with Exo70, a key regulator of exocytosis and actin dynamics, at the leading edge of migrating cells.
Hubert
Thomas
T
Department of Medical Protein Research, VIB, B-9000 Ghent, Belgium.
Vandekerckhove
Joël
J
Gettemans
Jan
J
D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology*; D004815:Epidermal Growth Factor / Q000235:genetics / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008856:Microscopy, Fluorescence; D028861:Nuclear Pore Complex Proteins; D011554:Pseudopodia / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014945:Wound Healing / Q000502:physiology
C547754:EXOC7 protein, human; D008562:Membrane Glycoproteins; D028861:Nuclear Pore Complex Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D033921:Vesicular Transport Proteins; C485184:enhanced green fluorescent protein; C049312:nuclear pore protein p62; D049452:Green Fluorescent Proteins; D004815:Epidermal Growth Factor