Home LiteratureArticle Details
PMID: 8707840 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Molecular and functional characterization of the p62 complex, an assembly of nuclear pore complex glycoproteins.

The Journal of cell biology ·Vol. 134 ·No. 3 ·1996-08-00 ·Pages 589-601

Hu T, Guan T, Gerace L

Abstract

Macromolecular trafficking across the nuclear envelope involves interactions between cytosolic transport factors and nuclear pore complex proteins. The p62 complex, an assembly of 62, 58, 54, and 45-kD O-linked glycoproteins-localized near the central gated channel of the nuclear pore complex, has been directly implicated in nuclear protein import. The cDNA cloning of rat p62 was reported previously. We have now carried out cDNA cloning of rat p58, p54, and p45. We found that p58 contains regions with FG (Phe, Gly) and PA (Pro, Ala) repeats at both its NH2 and COOH termini separated by a predicted alpha-helical coiled-coil region, while p54 has an NH2-terminal FG and PA repeat region and a COOH-terminal predicted coiled-coil region. p45 and p58 appear to be generated by alternative splicing, with p45 containing the NH2-terminal FG repeat region and the coiled-coil region of p58. Using immunogold electron microscopy, we found that p58/p45 and p54 are localized on both sides of the nuclear pore complex, like p62. Previous studies have shown that immobilized recombinant p62 can bind the cytosolic nuclear import factor NTF2 and thereby deplete transport activity from cytosol. We have now found that immobilized recombinant p58 and p54 also can deplete nuclear transport activity from cytosol, and that p62, p58, and p54 bind directly to the cytosolic nuclear import factors p97 and NTF2. At least in the case of p58, this involves FG repeat regions. Moreover, p58 can bind to a complex containing transport ligand, the nuclear localization sequence receptor (Srp1 alpha) and p97. These data support a model in which the p62 complex binds to a multicomponent particle consisting of transport ligand and cytosolic factors to achieve accumulation of ligand near the central gated channel of the nuclear pore complex.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Biological Transport Carrier Proteins/metabolism Cloning, Molecular Cytosol/metabolism DNA, Complementary/genetics HeLa Cells Humans Liver/chemistry Membrane Glycoproteins/analysis,chemistry,genetics,metabolism Membrane Proteins/metabolism Molecular Sequence Data Nuclear Envelope/chemistry Nuclear Pore Complex Proteins Nuclear Proteins/analysis,chemistry,genetics,metabolism Nucleocytoplasmic Transport Proteins Peptide Fragments/analysis,metabolism Protein Binding Protein Structure, Secondary Rats Recombinant Fusion Proteins alpha Karyopherins
Chemicals
Carrier Proteins DNA, Complementary Membrane Glycoproteins Membrane Proteins Nuclear Pore Complex Proteins Nuclear Proteins Nucleocytoplasmic Transport Proteins Nutf2 protein, rat Peptide Fragments Recombinant Fusion Proteins alpha Karyopherins nuclear pore protein p62 nuclear pore protein p97
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hu T
Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037, USA.
Guan T
Gerace L
References (55)
55 references, click to expand
  1. Cytosolic factors in nuclear transport: what's importin?
    Cell. 1994 Dec 16;79(6):931-4 PMID: 8001141
  2. Mammalian karyopherin alpha 1 beta and alpha 2 beta heterodimers: alpha 1 or alpha 2 subunit binds nuclear localization signal and beta subunit interacts with peptide repeat-containing nucleoporins.
    Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6532-6 PMID: 7604027
  3. Pores for thought: nuclear pore complex proteins.
    Trends Cell Biol. 1994 Oct;4(10):357-65 PMID: 14731624
  4. The GLFG repetitive region of the nucleoporin Nup116p interacts with Kap95p, an essential yeast nuclear import factor.
    J Cell Biol. 1995 Dec;131(6 Pt 2):1699-713 PMID: 8557738
  5. Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation.
    Cell. 1988 Mar 11;52(5):641-53 PMID: 3345567
  6. Sequence and characterization of cytoplasmic nuclear protein import factor p97.
    J Cell Biol. 1995 Jul;130(2):265-74 PMID: 7615630
  7. In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins.
    J Cell Biol. 1986 Dec;103(6 Pt 1):2091-102 PMID: 3097026
  8. Purification and characterization of a nuclear pore glycoprotein complex containing p62.
    J Biochem. 1993 Mar;113(3):377-82 PMID: 8486610
  9. 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
  10. 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
  11. Macromolecular interactions in the nucleoporin p62 complex of rat nuclear pores: binding of nucleoporin p54 to the rod domain of p62.
    J Cell Biol. 1995 Feb;128(3):251-61 PMID: 7531196
  12. Monoclonal antibodies identify a group of nuclear pore complex glycoproteins.
    J Cell Biol. 1987 May;104(5):1143-56 PMID: 2437126
  13. GTP hydrolysis by Ran occurs at the nuclear pore complex in an early step of protein import.
    J Cell Biol. 1995 Nov;131(3):571-81 PMID: 7593180
  14. Inhibition of nuclear protein import by nonhydrolyzable analogues of GTP and identification of the small GTPase Ran/TC4 as an essential transport factor.
    J Cell Biol. 1993 Dec;123(6 Pt 2):1649-59 PMID: 8276887
  15. Identification of a protein complex that is required for nuclear protein import and mediates docking of import substrate to distinct nucleoporins.
    Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1769-73 PMID: 7878057
  16. Previously identified protein of uncertain function is karyopherin alpha and together with karyopherin beta docks import substrate at nuclear pore complexes.
    Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2008-11 PMID: 7892216
  17. A short amino acid sequence able to specify nuclear location.
    Cell. 1984 Dec;39(3 Pt 2):499-509 PMID: 6096007
  18. Identification of hSRP1 alpha as a functional receptor for nuclear localization sequences.
    Science. 1995 May 19;268(5213):1049-53 PMID: 7754385
  19. A karyophilic protein forms a stable complex with cytoplasmic components prior to nuclear pore binding.
    J Biol Chem. 1995 Apr 14;270(15):8559-65 PMID: 7721756
  20. 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
  21. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence.
    Cell. 1991 Feb 8;64(3):615-23 PMID: 1991323
  22. A complex of nuclear pore proteins required for pore function.
    J Cell Biol. 1991 Jul;114(1):169-83 PMID: 2050741
  23. Structural analysis of the p62 complex, an assembly of O-linked glycoproteins that localizes near the central gated channel of the nuclear pore complex.
    Mol Biol Cell. 1995 Nov;6(11):1591-603 PMID: 8589458
  24. Nuclear transport.
    Curr Opin Cell Biol. 1994 Jun;6(3):335-42 PMID: 7917322
  25. Differential roles of heat shock protein 70 in the in vitro nuclear import of glucocorticoid receptor and simian virus 40 large tumor antigen.
    Mol Cell Biol. 1994 Aug;14(8):5088-98 PMID: 8035791
  26. Toward the molecular details of the nuclear pore complex.
    J Struct Biol. 1994 Nov-Dec;113(3):179-89 PMID: 7537513
  27. 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
  28. Antibodies against 70-kD heat shock cognate protein inhibit mediated nuclear import of karyophilic proteins.
    J Cell Biol. 1992 Dec;119(5):1047-61 PMID: 1332978
  29. RNA splicing: three themes with variations.
    Cell. 1983 Oct;34(3):713-6 PMID: 6354470
  30. Human nucleoporin p62 and the essential yeast nuclear pore protein NSP1 show sequence homology and a similar domain organization.
    Eur J Cell Biol. 1991 Jun;55(1):17-30 PMID: 1915414
  31. Identification of specific binding proteins for a nuclear location sequence.
    Nature. 1989 Jan 19;337(6204):276-9 PMID: 2911368
  32. In vivo evidence for involvement of a 58 kDa component of nuclear pore-targeting complex in nuclear protein import.
    EMBO J. 1995 Aug 1;14(15):3617-26 PMID: 7641681
  33. Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
    J Cell Biol. 1990 Sep;111(3):807-16 PMID: 2391365
  34. 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
  35. The human CAN protein, a putative oncogene product associated with myeloid leukemogenesis, is a nuclear pore complex protein that faces the cytoplasm.
    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1519-23 PMID: 8108440
  36. Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins.
    Anal Biochem. 1993 Apr;210(1):179-87 PMID: 8489015
  37. Mechanisms of nuclear protein import.
    Curr Opin Cell Biol. 1995 Jun;7(3):310-8 PMID: 7662359
  38. Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores.
    Cell. 1988 Mar 11;52(5):655-64 PMID: 3125984
  39. Two different subunits of importin cooperate to recognize nuclear localization signals and bind them to the nuclear envelope.
    Curr Biol. 1995 Apr 1;5(4):383-92 PMID: 7627554
  40. Primary sequence and heterologous expression of nuclear pore glycoprotein p62.
    J Cell Biol. 1990 Jun;110(6):1861-71 PMID: 2190987
  41. Nucleocytoplasmic transport.
    Science. 1996 Mar 15;271(5255):1513-8 PMID: 8599106
  42. Cytosolic proteins that specifically bind nuclear location signals are receptors for nuclear import.
    Cell. 1991 Sep 6;66(5):837-47 PMID: 1653647
  43. The transport of proteins into the nucleus requires the 70-kilodalton heat shock protein or its cytosolic cognate.
    Mol Cell Biol. 1992 May;12(5):2186-92 PMID: 1569948
  44. Identification of cytosolic factors required for nuclear location sequence-mediated binding to the nuclear envelope.
    J Cell Biol. 1994 May;125(3):547-55 PMID: 8175880
  45. Nuclear pore complex contains a family of glycoproteins that includes p62: glycosylation through a previously unidentified cellular pathway.
    Proc Natl Acad Sci U S A. 1987 Nov;84(21):7552-6 PMID: 3313397
  46. The importance of importin.
    Trends Cell Biol. 1995 May;5(5):189-91 PMID: 14731446
  47. Speculations on RNA splicing.
    Cell. 1981 Mar;23(3):643-6 PMID: 7226224
  48. Role of the nuclear transport factor p10 in nuclear import.
    Science. 1996 Apr 5;272(5258):120-2 PMID: 8600522
  49. 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
  50. The GTP-binding protein Ran/TC4 is required for protein import into the nucleus.
    Nature. 1993 Oct 14;365(6447):661-3 PMID: 8413630
  51. Cloning of a cDNA for lamina-associated polypeptide 2 (LAP2) and identification of regions that specify targeting to the nuclear envelope.
    EMBO J. 1995 Apr 18;14(8):1626-36 PMID: 7737115
  52. Organization and expression of eucaryotic split genes coding for proteins.
    Annu Rev Biochem. 1981;50:349-83 PMID: 6791577
  53. Isolation of a protein that is essential for the first step of nuclear protein import.
    Cell. 1994 Dec 2;79(5):767-78 PMID: 8001116
  54. Architecture and design of the nuclear pore complex.
    Cell. 1992 Jun 26;69(7):1133-41 PMID: 1617726
  55. Nuclear pore complex glycoprotein p62 of Xenopus laevis and mouse: cDNA cloning and identification of its glycosylated region.
    Eur J Cell Biol. 1991 Jun;55(1):31-47 PMID: 1915419
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.134.3.589
Published
1996-08-00
Pages
589-601
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120945
Subset
IM
Databases
GENBANK
AF000901, U63839, U63840
External Links
PubMed source DOI: 10.1083/jcb.134.3.589 PMC Full Text View journal details
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: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2