title: Size-dependent leak of soluble and membrane proteins through the yeast nuclear pore complex.
abstract: Nuclear pore complexes (NPCs) allow selective import and export while forming a barrier for untargeted proteins. Using fluorescence microscopy, we measured in vivo the permeability of the Saccharomyces cerevisiae NPC for multidomain proteins of different sizes and found that soluble proteins of 150 kDa and membrane proteins with an extralumenal domain of 90 kDa were still partly localized in the nucleus on a time scale of hours. The NPCs thus form only a weak barrier for the majority of yeast proteins, given their monomeric size. Using FGΔ-mutant strains, we showed that specific combinations of Nups, especially with Nup100, but not the total mass of FG-nups per pore, were important for forming the barrier. Models of the disordered phase of wild-type and mutant NPCs were generated using a one bead per amino acid molecular dynamics model. The permeability measurements correlated with the density predictions from coarse-grained molecular dynamics simulations in the center of the NPC. The combined in vivo and computational approach provides a framework for elucidating the structural and functional properties of the permeability barrier of nuclear pore complexes.
authors: European Research Institute for the Biology of Ageing, University of Groningen, University Medical Center Groningen, 9713 AV Groningen, Netherlands Zernike Institute for Advanced Materials and Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands.
authors: Petra
authors: P
authors: Popken
authors: Zernike Institute for Advanced Materials and.
authors: Ali
authors: A
authors: Ghavami
authors: Zernike Institute for Advanced Materials and.
authors: Patrick R
authors: PR
authors: Onck
authors: Zernike Institute for Advanced Materials and Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9747 AG Groningen, Netherlands.
authors: Bert
authors: B
authors: Poolman
authors: European Research Institute for the Biology of Ageing, University of Groningen, University Medical Center Groningen, 9713 AV Groningen, Netherlands l.m.veenhoff@rug.nl.
authors: Liesbeth M
authors: LM
authors: Veenhoff
mesh_terms: D004058:Diffusion; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D056004:Molecular Dynamics Simulation; D022022:Nuclear Pore / Q000378:metabolism*; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins
references: Science. 2011 Jul 1;333(6038):90-3
references: 21659568
references: EMBO Rep. 2007 Oct;8(10):914-9
references: 17906672
references: Nature. 2007 Nov 29;450(7170):695-701
references: 18046406
references: Proc Natl Acad Sci U S A. 2008 Apr 15;105(15):5722-7
references: 18391190
references: Mol Cell. 2008 Sep 26;31(6):925-32
references: 18922474
references: Cell. 2009 Jan 23;136(2):284-95
references: 19167330
references: EMBO J. 2009 Sep 2;28(17):2541-53
references: 19680228
references: EMBO J. 2009 Sep 16;28(18):2697-705
references: 19680225
references: Eukaryot Cell. 2009 Dec;8(12):1814-27
references: 19801417
references: PLoS Comput Biol. 2010 Apr;6(4):e1000747
references: 20421988
references: EMBO J. 2010 Jul 21;29(14):2262-75
references: 20551905
references: Trends Cell Biol. 2014 Apr;24(4):221-9
references: 24268652
references: Curr Opin Cell Biol. 2014 Jun;28:36-45
references: 24531277
references: Biophys J. 2014 Sep 16;107(6):1393-402
references: 25229147
references: J Cell Biol. 2000 Feb 21;148(4):635-51
references: 10684247
references: J Cell Biol. 2000 May 29;149(5):1027-38
references: 10831607
references: J Biol Chem. 2001 Nov 23;276(47):43548-56
references: 11557770
references: J Cell Sci. 2002 Apr 1;115(Pt 7):1361-71
references: 11896184
references: J Cell Sci. 2010 Aug 15;123(Pt 16):2773-80
references: 20647373
references: J Cell Biol. 2011 May 2;193(3):489-507
references: 21518795
references: Proc Natl Acad Sci U S A. 2012 May 8;109(19):7326-31
references: 22529346
references: Structure. 2012 Jun 6;20(6):998-1006
references: 22632834
references: Cell Mol Life Sci. 2012 Jul;69(13):2205-16
references: 22327555
references: Nat Methods. 2012 Jul;9(7):676-82
references: 22743772
references: Cell. 2012 Aug 17;150(4):738-51
references: 22901806
references: Cell. 2013 Mar 14;152(6):1218-21
references: 23498931
references: Traffic. 2013 May;14(5):487-501
references: 23357007
references: Nucleus. 2013 May-Jun;4(3):166-75
references: 23669120
references: Biophys J. 2013 Aug 6;105(3):565-9
references: 23931304
references: EMBO J. 2002 Jun 3;21(11):2664-71
references: 12032079
references: Mol Cell Biol. 2003 Jan;23(2):534-42
references: 12509452
references: Biophys J. 2003 Feb;84(2 Pt 1):1317-27
references: 12547812
references: Nature. 2003 Oct 16;425(6959):686-91
references: 14562095
references: Nat Cell Biol. 2004 Mar;6(3):197-206
references: 15039779
references: Biochem Biophys Res Commun. 2004 Jun 11;318(4):1079-84
references: 15147984
references: Cell. 1992 Jun 26;69(7):1133-41
references: 1617726
references: Genes Dev. 1992 Oct;6(10):1914-26
references: 1398069
references: Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):225-9
references: 7816822
references: Cell. 1995 Apr 21;81(2):215-22
references: 7736573
references: J Cell Biol. 1995 Jul;130(1):15-27
references: 7790369
references: J Cell Biol. 1995 Sep;130(5):1017-26
references: 7657689
references: Cell. 1995 Dec 1;83(5):683-92
references: 8521485
references: J Cell Biol. 1997 May 19;137(4):797-811
references: 9151683
references: J Cell Biol. 1997 Sep 22;138(6):1193-206
references: 9298976
references: Exp Cell Res. 1997 Oct 10;236(1):346-50
references: 9344616
references: J Cell Biol. 2004 Dec 20;167(6):1051-62
references: 15611332
references: Methods. 2006 Jan;38(1):17-24
references: 16343937
references: Nature. 2006 Aug 31;442(7106):1003-7
references: 16929305
references: Nature. 2006 Oct 19;443(7113):827-31
references: 17051211
references: Science. 2006 Nov 3;314(5800):815-7
references: 17082456
references: Cell. 2007 Apr 6;129(1):83-96
references: 17418788
references: FEBS Lett. 2007 Jul 10;581(17):3164-70
references: 17588566
references: Anal Biochem. 2007 Sep 1;368(1):95-9
references: 17586454
references: Proc Natl Acad Sci U S A. 2007 Jul 31;104(31):12743-8
references: 17646647
references: Genetics. 2012 Mar;190(3):855-83
references: 22419078