title: Defective nuclear import of Tpr in Progeria reflects the Ran sensitivity of large cargo transport.
abstract: The RanGTPase acts as a master regulator of nucleocytoplasmic transport by controlling assembly and disassembly of nuclear transport complexes. RanGTP is required in the nucleus to release nuclear localization signal (NLS)-containing cargo from import receptors, and, under steady-state conditions, Ran is highly concentrated in the nucleus. We previously showed the nuclear/cytoplasmic Ran distribution is disrupted in Hutchinson-Gilford Progeria syndrome (HGPS) fibroblasts that express the Progerin form of lamin A, causing a major defect in nuclear import of the protein, translocated promoter region (Tpr). In this paper, we show that Tpr import was mediated by the most abundant import receptor, KPNA2, which binds the bipartite NLS in Tpr with nanomolar affinity. Analyses including NLS swapping revealed Progerin did not cause global inhibition of nuclear import. Rather, Progerin inhibited Tpr import because transport of large protein cargoes was sensitive to changes in the Ran nuclear/cytoplasmic distribution that occurred in HGPS. We propose that defective import of large protein complexes with important roles in nuclear function may contribute to disease-associated phenotypes in Progeria.
authors: Center for Cell Signaling, University of Virginia, Charlottesville, VA 22903, USA.
authors: Chelsi J
authors: CJ
authors: Snow
authors: Ashraf
authors: A
authors: Dar
authors: Anindya
authors: A
authors: Dutta
authors: Ralph H
authors: RH
authors: Kehlenbach
authors: Bryce M
authors: BM
authors: Paschal
mesh_terms: D021581:Active Transport, Cell Nucleus; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D034904:Lamin Type A; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D010641:Phenotype; D011371:Progeria / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D028901:alpha Karyopherins / Q000235:genetics / Q000378:metabolism*; D020931:ran GTP-Binding Protein / Q000378:metabolism*
chemical_list: C490630:KPNA2 protein, human; D034904:Lamin Type A; D019913:Nuclear Localization Signals; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D011498:Protein Precursors; D011518:Proto-Oncogene Proteins; C073597:TPR protein, human; D028901:alpha Karyopherins; C090764:prelamin A; D020931:ran GTP-Binding Protein
references: Nucleic Acids Res. 2006 Jan 1;34(Database issue):D415-8
references: 16381901
references: Science. 2001 May 25;292(5521):1540-3
references: 11375490
references: Nucleus. 2012 Jul 1;3(4):349-58
references: 22688647
references: Proc Natl Acad Sci U S A. 2005 Sep 6;102(36):12879-84
references: 16129833
references: J Cell Biol. 2002 Oct 14;159(1):55-67
references: 12370244
references: Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):8963-8
references: 15184648
references: J Cell Sci. 2006 Nov 15;119(Pt 22):4644-9
references: 17062639
references: Mol Cell Biol. 1999 Nov;19(11):7782-91
references: 10523667
references: J Biol Chem. 1999 May 28;274(22):15820-7
references: 10336485
references: Nature. 2010 Sep 30;467(7315):600-3
references: 20811366
references: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8613-8
references: 18562297
references: Nucleic Acids Res. 2004 Jan 1;32(Database issue):D505-8
references: 14681468
references: Cold Spring Harb Perspect Biol. 2010 Oct;2(10):a000562
references: 20630994
references: EMBO J. 1998 Nov 16;17(22):6587-98
references: 9822603
references: EMBO J. 2010 May 19;29(10):1659-73
references: 20407419
references: Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3000-4
references: 1557405
references: J Cell Biol. 2003 Mar 31;160(7):1041-53
references: 12654904
references: Proc Natl Acad Sci U S A. 2006 Jun 6;103(23):8703-8
references: 16738054
references: Biochim Biophys Acta. 1994 Mar 1;1217(2):219-23
references: 8110839
references: Nature. 1991 Nov 7;354(6348):80-2
references: 1944575
references: Nature. 2003 May 15;423(6937):293-8
references: 12714972
references: J Cell Biol. 1998 Dec 28;143(7):1801-12
references: 9864356
references: Cold Spring Harb Perspect Biol. 2010 Feb;2(2):a000760
references: 20182615
references: Mol Cell. 2010 Jun 11;38(5):700-11
references: 20542002
references: J Biol Chem. 1997 Aug 22;272(34):21534-9
references: 9261173
references: Mol Biol Cell. 2007 Nov;18(11):4365-76
references: 17761537
references: Traffic. 2005 Mar;6(3):187-98
references: 15702987
references: Aging Cell. 2004 Aug;3(4):235-43
references: 15268757
references: Nat Rev Mol Cell Biol. 2010 Jul;11(7):490-501
references: 20571586
references: RNA. 2011 Jul;17(7):1344-56
references: 21613532
references: Cell Death Differ. 2007 Oct;14(10):1759-67
references: 17612587
references: Trends Cell Biol. 2008 Oct;18(10):456-66
references: 18786826
references: Biochem Cell Biol. 1999;77(4):321-9
references: 10546895
references: Cell. 1991 Feb 8;64(3):615-23
references: 1991323
references: J Biol Chem. 1996 Aug 2;271(31):18477-84
references: 8702493
references: Nat Rev Mol Cell Biol. 2002 Jul;3(7):487-97
references: 12094215
references: Cell. 1995 Dec 1;83(5):683-92
references: 8521485
references: J Biol Chem. 2010 Apr 2;285(14):10841-9
references: 20133940
references: Proc Natl Acad Sci U S A. 2009 Dec 8;106(49):20788-93
references: 19926845
references: Mol Biol Cell. 2004 Nov;15(11):4854-65
references: 15356261
references: FEBS Lett. 2008 Jun 18;582(14):2004-16
references: 18328825
references: Mol Biol Cell. 2003 May;14(5):1923-40
references: 12802065
references: Proc Natl Acad Sci U S A. 2005 Oct 4;102(40):14416-21
references: 16186497
references: FEBS Lett. 1989 Nov 6;257(2):411-4
references: 2583287
references: Genes Dev. 2008 Nov 1;22(21):2926-31
references: 18981471
references: PLoS One. 2012;7(1):e29921
references: 22253824
references: J Cell Biol. 2009 Mar 9;184(5):647-57
references: 19273613
references: J Cell Sci. 1994 Jan;107 ( Pt 1):61-7
references: 8175923
references: Curr Biol. 1998 Dec 17-31;8(25):1403-6
references: 9889103
references: Mol Biol Cell. 2004 Sep;15(9):4261-77
references: 15229283
references: Cell. 1997 Sep 19;90(6):1051-60
references: 9323133
references: Cell. 1997 Sep 19;90(6):1041-50
references: 9323132
references: EMBO J. 1996 Oct 15;15(20):5584-94
references: 8896452
references: Annu Rev Genomics Hum Genet. 2006;7:369-405
references: 16824021
references: BMC Cell Biol. 2010 Aug 11;11:63
references: 20701745
references: Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):4955-60
references: 17360326
references: Mol Cell Biol. 1999 Dec;19(12):8616-24
references: 10567585
references: Mol Biol Cell. 2000 Feb;11(2):703-19
references: 10679025
references: Exp Cell Res. 1998 Nov 25;245(1):43-56
references: 9828100
references: J Biol Chem. 2000 Jul 28;275(30):23175-80
references: 10811801
references: J Cell Biol. 1989 Oct;109(4 Pt 1):1389-97
references: 2677018
references: J Cell Sci. 1999 Jul;112 ( Pt 13):2253-64
references: 10362555
references: Mol Cell Biol. 2011 Aug;31(16):3378-95
references: 21670151
references: Mol Biol Cell. 2001 Aug;12(8):2433-52
references: 11514627
references: BMC Cell Biol. 2009 Oct 16;10:74
references: 19835572
references: J Cell Biol. 2008 Mar 10;180(5):857-65
references: 18316408
references: EMBO J. 2002 Jun 3;21(11):2664-71
references: 12032079
references: J Lipid Res. 2005 Dec;46(12):2531-58
references: 16207929