Home LiteratureArticle Details
PMID: 15703211 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Vertebrate Nup53 interacts with the nuclear lamina and is required for the assembly of a Nup93-containing complex.

Molecular biology of the cell ·Vol. 16 ·No. 5 ·2005-05-00 ·Pages 2382-94

Hawryluk-Gara LA, Shibuya EK, Wozniak RW

Abstract

The nuclear pore complex (NPC) is an evolutionarily conserved structure that mediates exchange of macromolecules across the nuclear envelope (NE). It is comprised of approximately 30 proteins termed nucleoporins that are each present in multiple copies. We have investigated the function of the human nucleoporin Nup53, the ortholog of Saccharomyces cerevisiae Nup53p. Both cell fractionation and in vitro binding data suggest that Nup53 is tightly associated with the NE membrane and the lamina where it interacts with lamin B. We have also shown that Nup53 is capable of physically interacting with a group of nucleoporins including Nup93, Nup155, and Nup205. Consistent with this observation, depletion of Nup53 using small interfering RNAs causes a decrease in the cellular levels of these nucleoporins as well as the spindle checkpoint protein Mad1, likely due to destabilization of Nup53-containing complexes. The cellular depletion of this group of nucleoporins, induced by depleting either Nup53 or Nup93, severely alters nuclear morphology producing phenotypes similar to that previously observed in cells depleted of lamin A and Mad1. On basis of these data, we propose a model in which Nup53 is positioned near the pore membrane and the lamina where it anchors an NPC subcomplex containing Nup93, Nup155, and Nup205.

MeSH Terms
Animals Cell Cycle Proteins/metabolism HeLa Cells Humans In Vitro Techniques Multiprotein Complexes Nuclear Lamina/metabolism Nuclear Pore/metabolism Nuclear Pore Complex Proteins/chemistry,genetics,metabolism Nuclear Proteins/metabolism Rats Recombinant Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Transfection Xenopus Proteins/chemistry,genetics,metabolism Xenopus laevis
Chemicals
Cell Cycle Proteins MAD1 protein, S cerevisiae MAD1L1 protein, human Multiprotein Complexes NUP53 protein, S cerevisiae Nuclear Pore Complex Proteins Nuclear Proteins Recombinant Proteins Saccharomyces cerevisiae Proteins Xenopus Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hawryluk-Gara Lisa A
Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
Shibuya Ellen K
Wozniak Richard W
References (65)
65 references, click to expand
  1. Nup155 is a novel nuclear pore complex protein that contains neither repetitive sequence motifs nor reacts with WGA.
    J Cell Biol. 1993 Apr;121(1):1-9 PMID: 8458861
  2. The RanGAP1-RanBP2 complex is essential for microtubule-kinetochore interactions in vivo.
    Curr Biol. 2004 Apr 6;14(7):611-7 PMID: 15062103
  3. Loss of A-type lamin expression compromises nuclear envelope integrity leading to muscular dystrophy.
    J Cell Biol. 1999 Nov 29;147(5):913-20 PMID: 10579712
  4. Structure and assembly of the Nup84p complex.
    J Cell Biol. 2000 Apr 3;149(1):41-54 PMID: 10747086
  5. The yeast nuclear pore complex: composition, architecture, and transport mechanism.
    J Cell Biol. 2000 Feb 21;148(4):635-51 PMID: 10684247
  6. The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm.
    Curr Opin Cell Biol. 2000 Jun;12(3):361-71 PMID: 10801463
  7. Stepwise reassembly of the nuclear envelope at the end of mitosis.
    J Cell Biol. 1993 Jul;122(2):295-306 PMID: 8391536
  8. Intranuclear filaments containing a nuclear pore complex protein.
    J Cell Biol. 1993 Dec;123(6 Pt 1):1333-44 PMID: 8253834
  9. Functional interaction of Nic96p with a core nucleoporin complex consisting of Nsp1p, Nup49p and a novel protein Nup57p.
    EMBO J. 1995 Jan 3;14(1):76-87 PMID: 7828598
  10. Two novel related yeast nucleoporins Nup170p and Nup157p: complementation with the vertebrate homologue Nup155p and functional interactions with the yeast nuclear pore-membrane protein Pom152p.
    J Cell Biol. 1995 Dec;131(5):1133-48 PMID: 8522578
  11. Nup120p: a yeast nucleoporin required for NPC distribution and mRNA transport.
    J Cell Biol. 1995 Dec;131(6 Pt 2):1659-75 PMID: 8557736
  12. A novel complex of nucleoporins, which includes Sec13p and a Sec13p homolog, is essential for normal nuclear pores.
    Cell. 1996 Jan 26;84(2):265-75 PMID: 8565072
  13. Nic96p is required for nuclear pore formation and functionally interacts with a novel nucleoporin, Nup188p.
    J Cell Biol. 1996 Jun;133(6):1141-52 PMID: 8682854
  14. The yeast nucleoporin Nup188p interacts genetically and physically with the core structures of the nuclear pore complex.
    J Cell Biol. 1996 Jun;133(6):1153-62 PMID: 8682855
  15. A novel ubiquitin-like modification modulates the partitioning of the Ran-GTPase-activating protein RanGAP1 between the cytosol and the nuclear pore complex.
    J Cell Biol. 1996 Dec;135(6 Pt 1):1457-70 PMID: 8978815
  16. Dynamics of nuclear pore distribution in nucleoporin mutant yeast cells.
    J Cell Biol. 1997 Feb 24;136(4):747-59 PMID: 9049242
  17. Nup93, a vertebrate homologue of yeast Nic96p, forms a complex with a novel 205-kDa protein and is required for correct nuclear pore assembly.
    Mol Biol Cell. 1997 Oct;8(10):2017-38 PMID: 9348540
  18. Three-dimensional architecture of the isolated yeast nuclear pore complex: functional and evolutionary implications.
    Mol Cell. 1998 Jan;1(2):223-34 PMID: 9659919
  19. Spindle checkpoint protein Xmad1 recruits Xmad2 to unattached kinetochores.
    J Cell Biol. 1998 Oct 19;143(2):283-95 PMID: 9786942
  20. Specific binding of the karyopherin Kap121p to a subunit of the nuclear pore complex containing Nup53p, Nup59p, and Nup170p.
    J Cell Biol. 1998 Dec 28;143(7):1813-30 PMID: 9864357
  21. A conserved biogenesis pathway for nucleoporins: proteolytic processing of a 186-kilodalton precursor generates Nup98 and the novel nucleoporin, Nup96.
    J Cell Biol. 1999 Mar 22;144(6):1097-112 PMID: 10087256
  22. Nup192p is a conserved nucleoporin with a preferential location at the inner site of the nuclear membrane.
    J Biol Chem. 1999 Aug 6;274(32):22646-51 PMID: 10428845
  23. Partial nuclear pore complex disassembly during closed mitosis in Aspergillus nidulans.
    Curr Biol. 2004 Nov 23;14(22):1973-84 PMID: 15556859
  24. Novel vertebrate nucleoporins Nup133 and Nup160 play a role in mRNA export.
    J Cell Biol. 2001 Oct 29;155(3):339-54 PMID: 11684705
  25. Transport into and out of the nucleus.
    Microbiol Mol Biol Rev. 2001 Dec;65(4):570-94, table of contents PMID: 11729264
  26. Identification of essential genes in cultured mammalian cells using small interfering RNAs.
    J Cell Sci. 2001 Dec;114(Pt 24):4557-65 PMID: 11792820
  27. The Mad2 spindle checkpoint protein undergoes similar major conformational changes upon binding to either Mad1 or Cdc20.
    Mol Cell. 2002 Jan;9(1):59-71 PMID: 11804586
  28. Modular self-assembly of a Y-shaped multiprotein complex from seven nucleoporins.
    EMBO J. 2002 Feb 1;21(3):387-97 PMID: 11823431
  29. SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles.
    J Cell Biol. 2002 Feb 18;156(4):595-602 PMID: 11854305
  30. Accelerating the rate of disassembly of karyopherin.cargo complexes.
    J Biol Chem. 2002 May 17;277(20):18161-72 PMID: 11867631
  31. The permeability barrier of nuclear pore complexes appears to operate via hydrophobic exclusion.
    EMBO J. 2002 Jun 3;21(11):2664-71 PMID: 12032079
  32. Proteomic analysis of the mammalian nuclear pore complex.
    J Cell Biol. 2002 Sep 2;158(5):915-27 PMID: 12196509
  33. Role of Hec1 in spindle checkpoint signaling and kinetochore recruitment of Mad1/Mad2.
    Science. 2002 Sep 27;297(5590):2267-70 PMID: 12351790
  34. Karyopherins in nuclear pore biogenesis: a role for Kap121p in the assembly of Nup53p into nuclear pore complexes.
    J Cell Biol. 2002 Oct 28;159(2):267-78 PMID: 12403813
  35. The yeast nuclear pore complex functionally interacts with components of the spindle assembly checkpoint.
    J Cell Biol. 2002 Dec 9;159(5):807-19 PMID: 12473689
  36. Deciphering networks of protein interactions at the nuclear pore complex.
    Mol Cell Proteomics. 2002 Dec;1(12):930-46 PMID: 12543930
  37. Depletion of a single nucleoporin, Nup107, prevents the assembly of a subset of nucleoporins into the nuclear pore complex.
    Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):981-5 PMID: 12552102
  38. Regulating access to the genome: nucleocytoplasmic transport throughout the cell cycle.
    Cell. 2003 Feb 21;112(4):441-51 PMID: 12600309
  39. The conserved Nup107-160 complex is critical for nuclear pore complex assembly.
    Cell. 2003 Apr 18;113(2):195-206 PMID: 12705868
  40. Removal of a single pore subcomplex results in vertebrate nuclei devoid of nuclear pores.
    Mol Cell. 2003 Apr;11(4):853-64 PMID: 12718872
  41. Nuclear membrane proteins with potential disease links found by subtractive proteomics.
    Science. 2003 Sep 5;301(5638):1380-2 PMID: 12958361
  42. Nup358 integrates nuclear envelope breakdown with kinetochore assembly.
    J Cell Biol. 2003 Sep 15;162(6):991-1001 PMID: 12963708
  43. The spindle assembly and spindle position checkpoints.
    Annu Rev Genet. 2003;37:251-82 PMID: 14616062
  44. Dynamic interactions of nuclear lamina proteins with chromatin and transcriptional machinery.
    Cell Mol Life Sci. 2003 Oct;60(10):2053-63 PMID: 14618255
  45. Virtual gating and nuclear transport: the hole picture.
    Trends Cell Biol. 2003 Dec;13(12):622-8 PMID: 14624840
  46. Caenorhabditis elegans nucleoporins Nup93 and Nup205 determine the limit of nuclear pore complex size exclusion in vivo.
    Mol Biol Cell. 2003 Dec;14(12):5104-15 PMID: 12937276
  47. Sorting out the nuclear envelope from the endoplasmic reticulum.
    Nat Rev Mol Cell Biol. 2004 Jan;5(1):65-9 PMID: 14663490
  48. The SONB(NUP98) nucleoporin interacts with the NIMA kinase in Aspergillus nidulans.
    Genetics. 2003 Nov;165(3):1071-81 PMID: 14668365
  49. Cell cycle regulated transport controlled by alterations in the nuclear pore complex.
    Cell. 2003 Dec 26;115(7):813-23 PMID: 14697200
  50. Yeast nucleoporins involved in passive nuclear envelope permeability.
    J Cell Biol. 2000 May 29;149(5):1027-38 PMID: 10831607
  51. Incorporation of the nuclear pore basket protein nup153 into nuclear pore structures is dependent upon lamina assembly: evidence from cell-free extracts of Xenopus eggs.
    EMBO J. 2000 Aug 1;19(15):3918-31 PMID: 10921874
  52. Identification of a new vertebrate nucleoporin, Nup188, with the use of a novel organelle trap assay.
    Mol Biol Cell. 2000 Oct;11(10):3381-96 PMID: 11029043
  53. The yeast nucleoporin Nup53p specifically interacts with Nic96p and is directly involved in nuclear protein import.
    Mol Biol Cell. 2000 Nov;11(11):3885-96 PMID: 11071914
  54. Mitotic checkpoint proteins HsMAD1 and HsMAD2 are associated with nuclear pore complexes in interphase.
    J Cell Sci. 2001 Mar;114(Pt 5):953-63 PMID: 11181178
  55. Kinetic analysis of translocation through nuclear pore complexes.
    EMBO J. 2001 Mar 15;20(6):1320-30 PMID: 11250898
  56. The karyopherin Kap142p/Msn5p mediates nuclear import and nuclear export of different cargo proteins.
    J Cell Biol. 2001 Feb 19;152(4):729-40 PMID: 11266464
  57. Lamins and apoptosis: a two-way street?
    J Cell Biol. 2001 Apr 30;153(3):F5-7 PMID: 11331313
  58. A link between the synthesis of nucleoporins and the biogenesis of the nuclear envelope.
    J Cell Biol. 2001 May 14;153(4):709-24 PMID: 11352933
  59. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells.
    Nature. 2001 May 24;411(6836):494-8 PMID: 11373684
  60. An evolutionarily conserved NPC subcomplex, which redistributes in part to kinetochores in mammalian cells.
    J Cell Biol. 2001 Sep 17;154(6):1147-60 PMID: 11564755
  61. The entire Nup107-160 complex, including three new members, is targeted as one entity to kinetochores in mitosis.
    Mol Biol Cell. 2004 Jul;15(7):3333-44 PMID: 15146057
  62. Nucleoporins as components of the nuclear pore complex core structure and Tpr as the architectural element of the nuclear basket.
    Mol Biol Cell. 2004 Sep;15(9):4261-77 PMID: 15229283
  63. Nuclei from rat liver: isolation method that combines purity with high yield.
    Science. 1966 Dec 30;154(3757):1662-5 PMID: 5924199
  64. Primary structure analysis of an integral membrane glycoprotein of the nuclear pore.
    J Cell Biol. 1989 Jun;108(6):2083-92 PMID: 2738089
  65. A nuclear pore complex protein that contains zinc finger motifs, binds DNA, and faces the nucleoplasm.
    Cell. 1993 Jan 15;72(1):29-38 PMID: 8422679
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
Published
2005-05-00
Epub
2005-00-09
Pages
2382-94
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1087243
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com