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PMID: 10747086 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Structure and assembly of the Nup84p complex.

The Journal of cell biology ·Vol. 149 ·No. 1 ·2000-04-03 ·Pages 41-54

Siniossoglou S, Lutzmann M, Santos-Rosa H, Leonard K, Mueller S, Aebi U, Hurt E

Abstract

The Nup84p complex consists of five nucleoporins (Nup84p, Nup85p, Nup120p, Nup145p-C, and Seh1p) and Sec13p, a bona fide subunit of the COPII coat complex. We show that a pool of green fluorescent protein-tagged Sec13p localizes to the nuclear pores in vivo, and identify sec13 mutant alleles that are synthetically lethal with nup85Delta and affect the localization of a green fluorescent protein-Nup49p reporter protein. In the electron microscope, sec13 mutants exhibit structural defects in nuclear pore complex (NPC) and nuclear envelope organization. For the assembly of the complex, Nup85p, Nup120p, and Nup145p-C are essential. A highly purified Nup84p complex was isolated from yeast under native conditions and its molecular mass was determined to be 375 kD by quantitative scanning transmission electron microscopy and analytical ultracentrifugation, consistent with a monomeric complex. Furthermore, the Nup84p complex exhibits a Y-shaped, triskelion-like morphology 25 nm in diameter in the transmission electron microscope. Thus, the Nup84p complex constitutes a paradigm of an NPC structural module with distinct composition, structure, and a role in nuclear mRNA export and NPC bio- genesis.

MeSH Terms
Alleles Amino Acid Sequence Cell Nucleus/chemistry,metabolism,ultrastructure Chromatography, Gel Epistasis, Genetic Fungal Proteins/genetics,isolation & purification,metabolism,ultrastructure Genes, Lethal/genetics Membrane Proteins/genetics,isolation & purification,metabolism,ultrastructure Microscopy, Electron, Scanning Transmission Molecular Sequence Data Molecular Weight Mutation/genetics Nuclear Envelope/chemistry,metabolism,ultrastructure Nuclear Pore Complex Proteins Nuclear Proteins/genetics,isolation & purification,metabolism,ultrastructure Porins/genetics,isolation & purification,metabolism,ultrastructure Protein Binding RNA-Binding Proteins/genetics,isolation & purification,metabolism,ultrastructure Recombinant Fusion Proteins/genetics,isolation & purification,metabolism,ultrastructure Saccharomyces cerevisiae/chemistry,cytology,genetics Saccharomyces cerevisiae Proteins Temperature Ultracentrifugation
Chemicals
Fungal Proteins Membrane Proteins NUP120 protein, S cerevisiae NUP145 protein, S cerevisiae NUP84 protein, S cerevisiae NUP85 protein, S cerevisiae Nuclear Pore Complex Proteins Nuclear Proteins Porins RNA-Binding Proteins Recombinant Fusion Proteins SEC13 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Siniossoglou S
Biochemie-Zentrum Heidelberg, D-69120 Heidelberg, Germany.
Lutzmann M
Santos-Rosa H
Leonard K
Mueller S
Aebi U
Hurt E
References (30)
30 references, click to expand
  1. Molecular weight determination by scanning transmission electron microscopy.
    Ultramicroscopy. 1978;3(3):273-81 PMID: 734784
  2. Dynamics of nuclear pore distribution in nucleoporin mutant yeast cells.
    J Cell Biol. 1997 Feb 24;136(4):747-59 PMID: 9049242
  3. Cytosolic Sec13p complex is required for vesicle formation from the endoplasmic reticulum in vitro.
    J Cell Biol. 1993 Feb;120(4):865-75 PMID: 8432727
  4. A temperature-sensitive NUP116 null mutant forms a nuclear envelope seal over the yeast nuclear pore complex thereby blocking nucleocytoplasmic traffic.
    J Cell Biol. 1993 Oct;123(2):275-84 PMID: 7691829
  5. The Sec13p complex and reconstitution of vesicle budding from the ER with purified cytosolic proteins.
    EMBO J. 1993 Nov;12(11):4073-82 PMID: 8223424
  6. Reversible random coil-beta-sheet transition of the Alzheimer beta-amyloid fragment (25-35).
    Biochemistry. 1994 Feb 15;33(6):1345-50 PMID: 8312252
  7. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum.
    Cell. 1994 Jun 17;77(6):895-907 PMID: 8004676
  8. Nup145p is required for nuclear export of mRNA and binds homopolymeric RNA in vitro via a novel conserved motif.
    Cell. 1994 Jul 29;78(2):275-89 PMID: 8044840
  9. Nup120p: a yeast nucleoporin required for NPC distribution and mRNA transport.
    J Cell Biol. 1995 Dec;131(6 Pt 2):1659-75 PMID: 8557736
  10. 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
  11. Pleiotropic nuclear defects associated with a conditional allele of the novel nucleoporin Rat9p/Nup85p.
    Mol Biol Cell. 1996 Jun;7(6):917-34 PMID: 8816998
  12. GLE2, a Saccharomyces cerevisiae homologue of the Schizosaccharomyces pombe export factor RAE1, is required for nuclear pore complex structure and function.
    Mol Biol Cell. 1996 Dec;7(12):1921-37 PMID: 8970155
  13. Dimples, pores, star-rings, and thin rings on growing nuclear envelopes: evidence for structural intermediates in nuclear pore complex assembly.
    J Cell Sci. 1997 Feb;110 ( Pt 4):409-20 PMID: 9067593
  14. From nucleoporins to nuclear pore complexes.
    Curr Opin Cell Biol. 1997 Jun;9(3):401-11 PMID: 9159086
  15. Mex67p, a novel factor for nuclear mRNA export, binds to both poly(A)+ RNA and nuclear pores.
    EMBO J. 1997 Jun 2;16(11):3256-71 PMID: 9214641
  16. Two functionally distinct domains generated by in vivo cleavage of Nup145p: a novel biogenesis pathway for nucleoporins.
    EMBO J. 1997 Aug 15;16(16):5086-97 PMID: 9305650
  17. Yeast genetics to dissect the nuclear pore complex and nucleocytoplasmic trafficking.
    Annu Rev Genet. 1997;31:277-313 PMID: 9442897
  18. Nup116p and nup100p are interchangeable through a conserved motif which constitutes a docking site for the mRNA transport factor gle2p.
    EMBO J. 1998 Feb 16;17(4):1107-19 PMID: 9463388
  19. Nucleocytoplasmic transport: the last 200 nanometers.
    Cell. 1998 Feb 6;92(3):327-36 PMID: 9476893
  20. Major binding sites for the nuclear import receptor are the internal nucleoporin Nup153 and the adjacent nuclear filament protein Tpr.
    J Cell Biol. 1998 Apr 6;141(1):31-49 PMID: 9531546
  21. Three-dimensional architecture of the isolated yeast nuclear pore complex: functional and evolutionary implications.
    Mol Cell. 1998 Jan;1(2):223-34 PMID: 9659919
  22. Transport from the endoplasmic reticulum to the Golgi.
    Curr Opin Cell Biol. 1998 Aug;10(4):477-82 PMID: 9719868
  23. A novel fluorescence-based genetic strategy identifies mutants of Saccharomyces cerevisiae defective for nuclear pore complex assembly.
    Mol Biol Cell. 1998 Sep;9(9):2439-61 PMID: 9725905
  24. Nuclear mRNA export requires complex formation between Mex67p and Mtr2p at the nuclear pores.
    Mol Cell Biol. 1998 Nov;18(11):6826-38 PMID: 9774696
  25. Molecular architecture of the yeast nuclear pore complex: localization of Nsp1p subcomplexes.
    J Cell Biol. 1998 Nov 2;143(3):577-88 PMID: 9813081
  26. A novel complex of membrane proteins required for formation of a spherical nucleus.
    EMBO J. 1998 Nov 16;17(22):6449-64 PMID: 9822591
  27. Proteins connecting the nuclear pore complex with the nuclear interior.
    J Cell Biol. 1999 Mar 8;144(5):839-55 PMID: 10085285
  28. 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
  29. The nuclear pore complex: from molecular architecture to functional dynamics.
    Curr Opin Cell Biol. 1999 Jun;11(3):391-401 PMID: 10395558
  30. Processing of quantitative scanning transmission electron micrographs.
    Scanning Microsc Suppl. 1988;2:285-93 PMID: 3244970
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.149.1.41
Published
2000-04-03
Pages
41-54
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2175103
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.149.1.41 PMC Full Text View journal details
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