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PMID: 11250898 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Kinetic analysis of translocation through nuclear pore complexes.

The EMBO journal ·Vol. 20 ·No. 6 ·2001-03-15 ·Pages 1320-30

Ribbeck K, Görlich D

Abstract

The mechanism of facilitated translocation through nuclear pore complexes (NPCs) is only poorly understood. Here, we present a kinetic analysis of the process using various model substrates. We find that the translocation capacity of NPCs is unexpectedly high, with a single NPC allowing a mass flow of nearly 100 MDa/s and rates in the order of 10(3) translocation events per second. Our data further indicate that high affinity interactions between the translocation substrate and NPC components are dispensable for translocation. We propose a 'selective phase model' that could explain how NPCs function as a permeability barrier for inert molecules and yet become selectively permeable for nuclear transport receptors and receptor-cargo complexes.

MeSH Terms
Active Transport, Cell Nucleus Carrier Proteins/metabolism Diffusion Green Fluorescent Proteins HeLa Cells Humans Karyopherins Luminescent Proteins/metabolism Models, Theoretical Nuclear Pore/metabolism Nuclear Proteins/metabolism Nucleocytoplasmic Transport Proteins Receptors, Cytoplasmic and Nuclear/metabolism
Chemicals
Carrier Proteins Karyopherins Luminescent Proteins Nuclear Proteins Nucleocytoplasmic Transport Proteins Receptors, Cytoplasmic and Nuclear Green Fluorescent Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ribbeck K
ZMBH, INF 282, 69120 Heidelberg, Germany.
Görlich D
References (49)
49 references, click to expand
  1. Three-dimensional architecture of the isolated yeast nuclear pore complex: functional and evolutionary implications.
    Mol Cell. 1998 Jan;1(2):223-34 PMID: 9659919
  2. Time sequence of nuclear pore formation in phytohemagglutinin-stimulated lymphocytes and in HeLa cells during the cell cycle.
    J Cell Biol. 1972 Nov;55(2):433-47 PMID: 5076782
  3. NTF2 mediates nuclear import of Ran.
    EMBO J. 1998 Nov 16;17(22):6587-98 PMID: 9822603
  4. Ran-dependent signal-mediated nuclear import does not require GTP hydrolysis by Ran.
    J Biol Chem. 1998 Dec 25;273(52):35170-5 PMID: 9857054
  5. Nuclear import of Ran is mediated by the transport factor NTF2.
    Curr Biol. 1998 Dec 17-31;8(25):1403-6 PMID: 9889103
  6. Receptor-mediated substrate translocation through the nuclear pore complex without nucleotide triphosphate hydrolysis.
    Curr Biol. 1999 Jan 14;9(1):30-41 PMID: 9889120
  7. The translocation of transportin-cargo complexes through nuclear pores is independent of both Ran and energy.
    Curr Biol. 1999 Jan 14;9(1):47-50 PMID: 9889126
  8. The importin beta/importin 7 heterodimer is a functional nuclear import receptor for histone H1.
    EMBO J. 1999 May 4;18(9):2411-23 PMID: 10228156
  9. Optical recording of signal-mediated protein transport through single nuclear pore complexes.
    Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):11842-7 PMID: 10518538
  10. Structural basis for molecular recognition between nuclear transport factor 2 (NTF2) and the GDP-bound form of the Ras-family GTPase Ran.
    J Mol Biol. 1998 Apr 3;277(3):635-46 PMID: 9533885
  11. Nuclear envelope permeability.
    Nature. 1975 Mar 13;254(5496):109-14 PMID: 1117994
  12. Movement of a karyophilic protein through the nuclear pores of oocytes.
    J Cell Biol. 1984 Dec;99(6):2216-22 PMID: 6501421
  13. Protein import through the nuclear pore complex is a multistep process.
    J Cell Biol. 1989 Sep;109(3):971-82 PMID: 2475512
  14. Accuracy and precision in the determination of Stokes radii and molecular masses of proteins by gel filtration chromatography.
    J Chromatogr. 1989 Jun 23;472(2):347-56 PMID: 2777944
  15. Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components.
    J Cell Biol. 1990 Apr;110(4):883-94 PMID: 2324201
  16. Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
    J Cell Biol. 1990 Sep;111(3):807-16 PMID: 2391365
  17. Catalysis of guanine nucleotide exchange on Ran by the mitotic regulator RCC1.
    Nature. 1991 Nov 7;354(6348):80-2 PMID: 1944575
  18. Architecture of the Xenopus nuclear pore complex revealed by three-dimensional cryo-electron microscopy.
    J Cell Biol. 1993 Jul;122(1):1-19 PMID: 8314837
  19. The GTP-binding protein Ran/TC4 is required for protein import into the nucleus.
    Nature. 1993 Oct 14;365(6447):661-3 PMID: 8413630
  20. Isolation of the yeast nuclear pore complex.
    J Cell Biol. 1993 Nov;123(4):771-83 PMID: 8227139
  21. Purification of a Ran-interacting protein that is required for protein import into the nucleus.
    Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):10212-6 PMID: 7937864
  22. The peptide repeat domain of nucleoporin Nup98 functions as a docking site in transport across the nuclear pore complex.
    Cell. 1995 Apr 21;81(2):215-22 PMID: 7736573
  23. Identification of NTF2, a cytosolic factor for nuclear import that interacts with nuclear pore complex protein p62.
    J Cell Biol. 1995 May;129(4):925-37 PMID: 7744965
  24. Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins.
    Cell. 1995 Dec 1;83(5):683-92 PMID: 8521485
  25. A novel receptor-mediated nuclear protein import pathway.
    Cell. 1996 Sep 20;86(6):985-94 PMID: 8808633
  26. Single nuclear pores visualized by confocal microscopy and image processing.
    Biophys J. 1996 May;70(5):2067-77 PMID: 9172731
  27. Identification of different roles for RanGDP and RanGTP in nuclear protein import.
    EMBO J. 1996 Oct 15;15(20):5584-94 PMID: 8896452
  28. Separate binding sites on nuclear transport factor 2 (NTF2) for GDP-Ran and the phenylalanine-rich repeat regions of nucleoporins p62 and Nsp1p.
    J Mol Biol. 1996 Nov 8;263(4):517-24 PMID: 8918934
  29. Dominant-negative mutants of importin-beta block multiple pathways of import and export through the nuclear pore complex.
    EMBO J. 1997 Mar 17;16(6):1153-63 PMID: 9135132
  30. Export of importin alpha from the nucleus is mediated by a specific nuclear transport factor.
    Cell. 1997 Sep 19;90(6):1061-71 PMID: 9323134
  31. Ran-unassisted nuclear migration of a 97-kD component of nuclear pore-targeting complex.
    J Cell Biol. 1997 Nov 17;139(4):841-9 PMID: 9362503
  32. RanBP1 is crucial for the release of RanGTP from importin beta-related nuclear transport factors.
    FEBS Lett. 1997 Dec 15;419(2-3):249-54 PMID: 9428644
  33. Import and export of the nuclear protein import receptor transportin by a mechanism independent of GTP hydrolysis.
    Curr Biol. 1998 Jan 15;8(2):89-95 PMID: 9427645
  34. Yeast genetics to dissect the nuclear pore complex and nucleocytoplasmic trafficking.
    Annu Rev Genet. 1997;31:277-313 PMID: 9442897
  35. Importins and exportins: how to get in and out of the nucleus.
    Trends Biochem Sci. 1998 May;23(5):185-9 PMID: 9612083
  36. The nuclear annuli as pathways for nucleocytoplasmic exchanges.
    J Cell Biol. 1962 Jul;14:65-72 PMID: 13892126
  37. Interaction between NTF2 and xFxFG-containing nucleoporins is required to mediate nuclear import of RanGDP.
    J Mol Biol. 1999 Oct 29;293(3):579-93 PMID: 10543952
  38. Transport between the cell nucleus and the cytoplasm.
    Annu Rev Cell Dev Biol. 1999;15:607-60 PMID: 10611974
  39. Transport of proteins and RNAs in and out of the nucleus.
    Cell. 1999 Dec 23;99(7):677-90 PMID: 10619422
  40. The C-terminal domain of TAP interacts with the nuclear pore complex and promotes export of specific CTE-bearing RNA substrates.
    RNA. 2000 Jan;6(1):136-58 PMID: 10668806
  41. Mapping interactions between nuclear transport factors in living cells reveals pathways through the nuclear pore complex.
    Mol Cell. 2000 Jan;5(1):133-40 PMID: 10678175
  42. Dissecting the interactions between NTF2, RanGDP, and the nucleoporin XFXFG repeats.
    J Biol Chem. 2000 Feb 25;275(8):5874-9 PMID: 10681579
  43. The yeast nuclear pore complex: composition, architecture, and transport mechanism.
    J Cell Biol. 2000 Feb 21;148(4):635-51 PMID: 10684247
  44. Anomalous diffusion of fluorescent probes inside living cell nuclei investigated by spatially-resolved fluorescence correlation spectroscopy.
    J Mol Biol. 2000 May 12;298(4):677-89 PMID: 10788329
  45. The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm.
    Curr Opin Cell Biol. 2000 Jun;12(3):361-71 PMID: 10801463
  46. Nuclear import of the ran exchange factor, RCC1, is mediated by at least two distinct mechanisms.
    J Cell Biol. 2000 May 15;149(4):835-50 PMID: 10811825
  47. Structural basis for the interaction between FxFG nucleoporin repeats and importin-beta in nuclear trafficking.
    Cell. 2000 Jul 7;102(1):99-108 PMID: 10929717
  48. Binding of the Mex67p/Mtr2p heterodimer to FXFG, GLFG, and FG repeat nucleoporins is essential for nuclear mRNA export.
    J Cell Biol. 2000 Aug 21;150(4):695-706 PMID: 10952996
  49. Nucleocytoplasmic transport: the soluble phase.
    Annu Rev Biochem. 1998;67:265-306 PMID: 9759490
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.6.1320
Published
2001-03-15
Pages
1320-30
Language
English
Region
England
NLM ID
8208664
PMCID
PMC145537
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.6.1320 PMC Full Text View journal details
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