Europe PMC

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Nup120p: a yeast nucleoporin required for NPC distribution and mRNA transport.

Author information

Affiliations

  • 1. Laboratory of Cell Biology, Howard Hughes Medical Institute, Rockefeller University, New York 10021-6399, USA.
      Authors
    • Aitchison JD 1
    (1 author)

ORCIDs linked to this article

The Journal of Cell Biology, 01 Dec 1995, 131(6 Pt 2):1659-1675
https://doi.org/10.1083/jcb.131.6.1659 PMID: 8557736 PMCID: PMC2120676

This article is in the Europe PMC Open access subset. Refer to the copyright information in the article for licensing details.
Free full text in Europe PMC

Abstract 

To extend our understanding of the mechanism by which the nuclear pore complex (NPC) mediates macromolecular transport across the nuclear envelope we have focused on defining the composition and molecular organization of the yeast NPC. Peptide sequence analysis of a polypeptide with a M(r) of approximately 100,000 present in a highly enriched yeast NPC fraction identified a novel yeast nucleoporin we term Nup120p. Nup120p corresponds to the open reading frame (ORF) YKL057c identified by the yeast genome sequencing project. The ORF predicts a protein with a calculated molecular mass of 120.5 kD containing two leucine zipper motifs, a short coiled-coil region and limited primary sequence similarity to Nup133p. Nup120p was localized to the NPC using a protein A-tagged chimera in situ by indirect immunofluorescence microscopy. Deletion of the NUP120 gene caused clustering of NPCs at one side of the nuclear envelope, moderate nucleolar fragmentation and slower cell growth. Transfer of nup120 delta cells to 37 degrees C resulted in the nuclear accumulation of poly(A)+ mRNA, extensive fragmentation of the nucleolus, spindle defects, and cell death.

Free full text 

Logo of jcellbiolLink to Publisher's site
J Cell Biol. 1995 Dec 2; 131(6): 1659–1675.
PMCID: PMC2120676
PMID: 8557736

Nup120p: a yeast nucleoporin required for NPC distribution and mRNA transport

Copyright and License information Disclaimer

Abstract

To extend our understanding of the mechanism by which the nuclear pore complex (NPC) mediates macromolecular transport across the nuclear envelope we have focused on defining the composition and molecular organization of the yeast NPC. Peptide sequence analysis of a polypeptide with a M(r) of approximately 100,000 present in a highly enriched yeast NPC fraction identified a novel yeast nucleoporin we term Nup120p. Nup120p corresponds to the open reading frame (ORF) YKL057c identified by the yeast genome sequencing project. The ORF predicts a protein with a calculated molecular mass of 120.5 kD containing two leucine zipper motifs, a short coiled-coil region and limited primary sequence similarity to Nup133p. Nup120p was localized to the NPC using a protein A-tagged chimera in situ by indirect immunofluorescence microscopy. Deletion of the NUP120 gene caused clustering of NPCs at one side of the nuclear envelope, moderate nucleolar fragmentation and slower cell growth. Transfer of nup120 delta cells to 37 degrees C resulted in the nuclear accumulation of poly(A)+ mRNA, extensive fragmentation of the nucleolus, spindle defects, and cell death.

Full Text

The Full Text of this article is available as a PDF (5.6M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aitchison JD, Rout MP, Marelli M, Blobel G, Wozniak RW. 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):1133–1148. [Europe PMC free article] [Abstract] [Google Scholar]
  • Akey CW. Interactions and structure of the nuclear pore complex revealed by cryo-electron microscopy. J Cell Biol. 1989 Sep;109(3):955–970. [Europe PMC free article] [Abstract] [Google Scholar]
  • Akey CW, Radermacher M. Architecture of the Xenopus nuclear pore complex revealed by three-dimensional cryo-electron microscopy. J Cell Biol. 1993 Jul;122(1):1–19. [Europe PMC free article] [Abstract] [Google Scholar]
  • Alber T. Structure of the leucine zipper. Curr Opin Genet Dev. 1992 Apr;2(2):205–210. [Abstract] [Google Scholar]
  • Aris JP, Blobel G. Identification and characterization of a yeast nucleolar protein that is similar to a rat liver nucleolar protein. J Cell Biol. 1988 Jul;107(1):17–31. [Europe PMC free article] [Abstract] [Google Scholar]
  • Belanger KD, Kenna MA, Wei S, Davis LI. Genetic and physical interactions between Srp1p and nuclear pore complex proteins Nup1p and Nup2p. J Cell Biol. 1994 Aug;126(3):619–630. [Europe PMC free article] [Abstract] [Google Scholar]
  • Byers B, Goetsch L. Behavior of spindles and spindle plaques in the cell cycle and conjugation of Saccharomyces cerevisiae. J Bacteriol. 1975 Oct;124(1):511–523. [Europe PMC free article] [Abstract] [Google Scholar]
  • Byrd DA, Sweet DJ, Panté N, Konstantinov KN, Guan T, Saphire AC, Mitchell PJ, Cooper CS, Aebi U, Gerace L. Tpr, a large coiled coil protein whose amino terminus is involved in activation of oncogenic kinases, is localized to the cytoplasmic surface of the nuclear pore complex. J Cell Biol. 1994 Dec;127(6 Pt 1):1515–1526. [Europe PMC free article] [Abstract] [Google Scholar]
  • Delorme E. Transformation of Saccharomyces cerevisiae by electroporation. Appl Environ Microbiol. 1989 Sep;55(9):2242–2246. [Europe PMC free article] [Abstract] [Google Scholar]
  • Doye V, Wepf R, Hurt EC. A novel nuclear pore protein Nup133p with distinct roles in poly(A)+ RNA transport and nuclear pore distribution. EMBO J. 1994 Dec 15;13(24):6062–6075. [Europe PMC free article] [Abstract] [Google Scholar]
  • Enenkel C, Blobel G, Rexach M. Identification of a yeast karyopherin heterodimer that targets import substrate to mammalian nuclear pore complexes. J Biol Chem. 1995 Jul 14;270(28):16499–16502. [Abstract] [Google Scholar]
  • Fabre E, Hurt EC. Nuclear transport. Curr Opin Cell Biol. 1994 Jun;6(3):335–342. [Abstract] [Google Scholar]
  • Fernandez J, Andrews L, Mische SM. An improved procedure for enzymatic digestion of polyvinylidene difluoride-bound proteins for internal sequence analysis. Anal Biochem. 1994 Apr;218(1):112–117. [Abstract] [Google Scholar]
  • Forrester W, Stutz F, Rosbash M, Wickens M. Defects in mRNA 3'-end formation, transcription initiation, and mRNA transport associated with the yeast mutation prp20: possible coupling of mRNA processing and chromatin structure. Genes Dev. 1992 Oct;6(10):1914–1926. [Abstract] [Google Scholar]
  • Fuchs R. MacPattern: protein pattern searching on the Apple Macintosh. Comput Appl Biosci. 1991 Jan;7(1):105–106. [Abstract] [Google Scholar]
  • Gerace L, Ottaviano Y, Kondor-Koch C. Identification of a major polypeptide of the nuclear pore complex. J Cell Biol. 1982 Dec;95(3):826–837. [Europe PMC free article] [Abstract] [Google Scholar]
  • Goldberg MW, Blow JJ, Allen TD. The use of field emission in-lens scanning electron microscopy to study the steps of assembly of the nuclear envelope in vitro. J Struct Biol. 1992 May-Jun;108(3):257–268. [Abstract] [Google Scholar]
  • Görlich D, Vogel F, Mills AD, Hartmann E, Laskey RA. Distinct functions for the two importin subunits in nuclear protein import. Nature. 1995 Sep 21;377(6546):246–248. [Abstract] [Google Scholar]
  • Gorsch LC, Dockendorff TC, Cole CN. A conditional allele of the novel repeat-containing yeast nucleoporin RAT7/NUP159 causes both rapid cessation of mRNA export and reversible clustering of nuclear pore complexes. J Cell Biol. 1995 May;129(4):939–955. [Europe PMC free article] [Abstract] [Google Scholar]
  • Grandi P, Doye V, Hurt EC. Purification of NSP1 reveals complex formation with 'GLFG' nucleoporins and a novel nuclear pore protein NIC96. EMBO J. 1993 Aug;12(8):3061–3071. [Europe PMC free article] [Abstract] [Google Scholar]
  • Grandi P, Schlaich N, Tekotte H, Hurt EC. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hallberg E, Wozniak RW, Blobel G. An integral membrane protein of the pore membrane domain of the nuclear envelope contains a nucleoporin-like region. J Cell Biol. 1993 Aug;122(3):513–521. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hinshaw JE, Carragher BO, Milligan RA. Architecture and design of the nuclear pore complex. Cell. 1992 Jun 26;69(7):1133–1141. [Abstract] [Google Scholar]
  • Hurt EC. A novel nucleoskeletal-like protein located at the nuclear periphery is required for the life cycle of Saccharomyces cerevisiae. EMBO J. 1988 Dec 20;7(13):4323–4334. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hurwitz ME, Blobel G. NUP82 is an essential yeast nucleoporin required for poly(A)+ RNA export. J Cell Biol. 1995 Sep;130(6):1275–1281. [Europe PMC free article] [Abstract] [Google Scholar]
  • Jarnik M, Aebi U. Toward a more complete 3-D structure of the nuclear pore complex. J Struct Biol. 1991 Dec;107(3):291–308. [Abstract] [Google Scholar]
  • Kadowaki T, Hitomi M, Chen S, Tartakoff AM. Nuclear mRNA accumulation causes nucleolar fragmentation in yeast mtr2 mutant. Mol Biol Cell. 1994 Nov;5(11):1253–1263. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kilmartin JV, Adams AE. Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces. J Cell Biol. 1984 Mar;98(3):922–933. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kraemer DM, Strambio-de-Castillia C, Blobel G, Rout MP. The essential yeast nucleoporin NUP159 is located on the cytoplasmic side of the nuclear pore complex and serves in karyopherin-mediated binding of transport substrate. J Biol Chem. 1995 Aug 11;270(32):19017–19021. [Abstract] [Google Scholar]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [Abstract] [Google Scholar]
  • Lee WC, Xue ZX, Mélèse T. The NSR1 gene encodes a protein that specifically binds nuclear localization sequences and has two RNA recognition motifs. J Cell Biol. 1991 Apr;113(1):1–12. [Europe PMC free article] [Abstract] [Google Scholar]
  • Li O, Heath CV, Amberg DC, Dockendorff TC, Copeland CS, Snyder M, Cole CN. Mutation or deletion of the Saccharomyces cerevisiae RAT3/NUP133 gene causes temperature-dependent nuclear accumulation of poly(A)+ RNA and constitutive clustering of nuclear pore complexes. Mol Biol Cell. 1995 Apr;6(4):401–417. [Europe PMC free article] [Abstract] [Google Scholar]
  • Lipman DJ, Pearson WR. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. [Abstract] [Google Scholar]
  • Loeb JD, Schlenstedt G, Pellman D, Kornitzer D, Silver PA, Fink GR. The yeast nuclear import receptor is required for mitosis. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7647–7651. [Europe PMC free article] [Abstract] [Google Scholar]
  • Maul GG. The nuclear and the cytoplasmic pore complex: structure, dynamics, distribution, and evolution. Int Rev Cytol Suppl. 1977;(6):75–186. [Abstract] [Google Scholar]
  • Moroianu J, Blobel G, Radu A. 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):2008–2011. [Europe PMC free article] [Abstract] [Google Scholar]
  • Moroianu J, Hijikata M, Blobel G, Radu A. 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):6532–6536. [Europe PMC free article] [Abstract] [Google Scholar]
  • Nehrbass U, Kern H, Mutvei A, Horstmann H, Marshallsay B, Hurt EC. NSP1: a yeast nuclear envelope protein localized at the nuclear pores exerts its essential function by its carboxy-terminal domain. Cell. 1990 Jun 15;61(6):979–989. [Abstract] [Google Scholar]
  • Pemberton LF, Rout MP, Blobel G. Disruption of the nucleoporin gene NUP133 results in clustering of nuclear pore complexes. Proc Natl Acad Sci U S A. 1995 Feb 14;92(4):1187–1191. [Europe PMC free article] [Abstract] [Google Scholar]
  • Radu A, Blobel G, Wozniak RW. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Radu A, Blobel G, Wozniak RW. Nup107 is a novel nuclear pore complex protein that contains a leucine zipper. J Biol Chem. 1994 Jul 1;269(26):17600–17605. [Abstract] [Google Scholar]
  • Radu A, Blobel G, Moore MS. 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):1769–1773. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ris H, Malecki M. High-resolution field emission scanning electron microscope imaging of internal cell structures after Epon extraction from sections: a new approach to correlative ultrastructural and immunocytochemical studies. J Struct Biol. 1993 Sep-Oct;111(2):148–157. [Abstract] [Google Scholar]
  • Rose M, Grisafi P, Botstein D. Structure and function of the yeast URA3 gene: expression in Escherichia coli. Gene. 1984 Jul-Aug;29(1-2):113–124. [Abstract] [Google Scholar]
  • Rothstein R. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol. 1991;194:281–301. [Abstract] [Google Scholar]
  • Rout MP, Kilmartin JV. Components of the yeast spindle and spindle pole body. J Cell Biol. 1990 Nov;111(5 Pt 1):1913–1927. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rout MP, Blobel G. Isolation of the yeast nuclear pore complex. J Cell Biol. 1993 Nov;123(4):771–783. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rout MP, Wente SR. Pores for thought: nuclear pore complex proteins. Trends Cell Biol. 1994 Oct;4(10):357–365. [Abstract] [Google Scholar]
  • Schneiter R, Kadowaki T, Tartakoff AM. mRNA transport in yeast: time to reinvestigate the functions of the nucleolus. Mol Biol Cell. 1995 Apr;6(4):357–370. [Europe PMC free article] [Abstract] [Google Scholar]
  • Stutz F, Neville M, Rosbash M. Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast. Cell. 1995 Aug 11;82(3):495–506. [Abstract] [Google Scholar]
  • Uhlén M, Guss B, Nilsson B, Gatenbeck S, Philipson L, Lindberg M. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications. J Biol Chem. 1984 Feb 10;259(3):1695–1702. [Abstract] [Google Scholar]
  • Unwin PN, Milligan RA. A large particle associated with the perimeter of the nuclear pore complex. J Cell Biol. 1982 Apr;93(1):63–75. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wente SR, Blobel G. 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):275–284. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wente SR, Blobel G. NUP145 encodes a novel yeast glycine-leucine-phenylalanine-glycine (GLFG) nucleoporin required for nuclear envelope structure. J Cell Biol. 1994 Jun;125(5):955–969. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wente SR, Rout MP, Blobel G. A new family of yeast nuclear pore complex proteins. J Cell Biol. 1992 Nov;119(4):705–723. [Europe PMC free article] [Abstract] [Google Scholar]
  • Winey M, Mamay CL, O'Toole ET, Mastronarde DN, Giddings TH, Jr, McDonald KL, McIntosh JR. Three-dimensional ultrastructural analysis of the Saccharomyces cerevisiae mitotic spindle. J Cell Biol. 1995 Jun;129(6):1601–1615. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wozniak RW, Bartnik E, Blobel G. Primary structure analysis of an integral membrane glycoprotein of the nuclear pore. J Cell Biol. 1989 Jun;108(6):2083–2092. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wozniak RW, Blobel G, Rout MP. POM152 is an integral protein of the pore membrane domain of the yeast nuclear envelope. J Cell Biol. 1994 Apr;125(1):31–42. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wu J, Matunis MJ, Kraemer D, Blobel G, Coutavas E. Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region. J Biol Chem. 1995 Jun 9;270(23):14209–14213. [Abstract] [Google Scholar]
  • Yang CH, Lambie EJ, Hardin J, Craft J, Snyder M. Higher order structure is present in the yeast nucleus: autoantibody probes demonstrate that the nucleolus lies opposite the spindle pole body. Chromosoma. 1989 Aug;98(2):123–128. [Abstract] [Google Scholar]
  • Yano R, Oakes M, Yamaghishi M, Dodd JA, Nomura M. Cloning and characterization of SRP1, a suppressor of temperature-sensitive RNA polymerase I mutations, in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Dec;12(12):5640–5651. [Europe PMC free article] [Abstract] [Google Scholar]
Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

Full text links 

Read article at publisher's site: https://doi.org/10.1083/jcb.131.6.1659

Read article for free, from open access legal sources, via Unpaywall: https://rupress.org/jcb/article-pdf/131/6/1659/1265193/1659.pdfUnpaywall

Citations & impact 

Impact metrics

113
Citations
Jump to Citations
5
Data citations
Jump to Data

Citations of article over time

Smart citations by scite.ai
Smart citations by scite.ai include citation statements extracted from the full text of the citing article. The number of the statements may be higher than the number of citations provided by EuropePMC if one paper cites another multiple times or lower if scite has not yet processed some of the citing articles.
Explore citation contexts and check if this article has been supported or disputed.
https://scite.ai/reports/10.1083/jcb.131.6.1659

Supporting
Mentioning
Contrasting
8
155
2

Article citations

Go to all (113) article citations

Data 

Similar Articles 

To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation.