Advertisement

Antonie van Leeuwenhoek

, Volume 62, Issue 1–2, pp 3–14 | Cite as

Nuclear transport and nuclear pores in yeast

  • U. Nehrbass
  • E. C. Hurt
Article

Abstract

The central features of nuclear import have been conserved during evolution. In yeast the nuclear accumulation of proteins follows the same selective and active transport mechanisms known from higher eukaryotes. Yeast nuclear proteins contain nuclear localization sequences (NLS) which are presumably recognized by receptors in the cytoplasm and the nuclear envelope. Subsequent to this recognition step, nuclear proteins are translocated into the nucleus via the nuclear pore complexes. The structure of the yeast nuclear pore complex resembles that of higher eukaryotes. Recently, the first putative components of the yeast nuclear import machinery have been cloned and sequenced. The genetically amenable yeast system allows for an efficient structural and functional analysis of these components. Due to the evolutionary conservation potential insights into the nuclear import mechanisms in yeast can be transferred to higher eukaryotes. Thus, yeast can be considered as a eukaryotic model system to study nuclear transport.

Key words

Saccharomyces cerevisiae nuclear pore complex nuclear transport nuclear localization sequence nucleoporins 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adam SA & Gerace L (1991) Cytosolic proteins that specifically bind nuclear localization signals are receptors for nuclear import. Cell 66: 837–847.Google Scholar
  2. Akey CW (1989) Interactions and structure of the nuclear pore complex revealed by cryo-electron microscopy. J. Cell Biol. 109: 955–970Google Scholar
  3. (1990) Visualization of transport-related configurations of the nuclear pore transporter. Biophys. J. 58: 341–355Google Scholar
  4. Akey CW & Goldfarb DS (1989) Protein import through the nuclear pore complex is a multistep process. J. Cell Biol. 109: 971–982Google Scholar
  5. Allen JL & Douglas MG (1989) Organization of the nuclear pore complex inSaccharomyces cerevisiae. J. Ultrastruc. Molec. Struct. Res. 102: 95–108Google Scholar
  6. Amati BB & Gasser SM (1988) Chromosomal ARS and CEN elements bind specifically to the yeast nuclear scaffold. Cell 54: 967–978Google Scholar
  7. Aris JP & Blobel G (1988) Identification and characterization of a yeast nucleolar protein that is similar to a rat liver nucleolar protein. J. Cell Biol. 107: 17–31.Google Scholar
  8. (1989) Yeast nuclear envelope proteins cross-react with an antibody against mammalian pore complex proteins. J. Cell Biol. 108: 2059–2067Google Scholar
  9. Bäuerele PA & Baltimore D (1988) IϰB: A specific inhibitor of NF-ϰB transription factor. Science 242: 540–546Google Scholar
  10. Barnes G & Rine J (1985) Regulated expression of endonuclease EcoRI inSaccharomyces cerevisiae: nuclear entry and biological consequences. Proc. Natl. Acad. Sci. USA. 82: 1354–1358Google Scholar
  11. Bataillé N, Helser T & Fried HM (1990) Cytoplasmic transport of ribosomal subunits microinjected into theXenopus laevis oocyte nucleus: a generalized, facilitated process. J. Cell Biol. 111: 1571–1582Google Scholar
  12. Breeuwer M & Goldfarb M (1990) Facilitated nuclear transport of histone H1 and other small nucleophilic proteins. Cell 60: 999–1008Google Scholar
  13. Cardenas ME, Laroche T & Gasser SM (1990) The composition and morphology of yeast nuclear scaffolds. J. Cell Sci. 96: 439–450Google Scholar
  14. Carmo-Fonseca M, Cidadao AJ & David-Ferreira JF (1987) Filamentous cross-bridges link intermediate filaments to the nuclear pore complexes. Eur. J. Cell Biol. 45: 282–290Google Scholar
  15. Carmo-Fonseca M, Kern H & Hurt EC (1991) Human nucleoporin p62 and the essential yeast nuclear pore protein NSP1 show sequence homology and a similar domain organization. Eur. J. Cell Biol. 55: 17–30Google Scholar
  16. Cohen C & Parry DAD (1990) α-helical coiled coils and bundles: how to design an α-helical protein. Proteins 7: 1–15Google Scholar
  17. Cordes V, Waizenegger I & Krohne G (1991) Nuclear pore complex glycoprotein p62 ofXenopus laevis and mouse: cDNA cloning and identification of its glycosylated region. Eur. J. Cell Biol. 55: 31–47Google Scholar
  18. Dabauvalle MC, Schulz B, Scheer U & Peters R (1988) Inhibition of nuclear accumulation of karyophilic proteins in living cells by microinjection of the lectin wheat germ agglutinin. Exp. Cell Res. 174: 291–296Google Scholar
  19. Dabauvalle M-C, Loos K & Scheer U (1990) Identification of a soluble precursor complex essential for nuclear pore assembly. Chromosoma 100: 56–66Google Scholar
  20. Davis LI & Blobel (1986) Identification and characterization of a nuclear pore complex protein. Cell 45: 699–709Google Scholar
  21. (1987) Nuclear pore complex contains a family of glycoproteins that includes p62: glycosylation through a previously unidentified cellular pathway. Proc. Natl. Acad. Sci. USA 84: 7552–7556Google Scholar
  22. Davis LI & Fink GR (1990) The NUP1 gene encodes an essential component of the yeast nuclear pore complex. Cell 61: 965–978Google Scholar
  23. Dingwall C, Sharnick SV & Laskey RA (1982) A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell 30: 449–458Google Scholar
  24. Featherstone C, Darby MK & Gerace L (1988) A monoclonal antibody against the nuclear pore complex inhibits nucleocytoplasmic transport of protein and RNAin-vivo. J. Cell Biol. 107: 1289–1297Google Scholar
  25. Feldherr CM, Kallenbach E & Schultz N (1984) Movement of a karyophilic protein through the nuclear pores of oocytes. J. Cell Biol. 99: 2216–2222Google Scholar
  26. Finlay DR & Forbes DJ (1990) Reconstitution of biochemically altered nuclear pores: transport can be eliminated and restored. Cell 60: 17–29Google Scholar
  27. Finlay DR, Newmeyer DD, Price TM & Forbes DJ (1987) Inhibition ofin-vitro nuclear transport by a lectin that binds to nuclear pores. J. Cell Biol. 104: 189–200Google Scholar
  28. Finlay DR, Meier E, Bradley P, Horecka J & Forbes DJ (1991) A complex of nuclear pore proteins required for pore functions. J. Cell Biol. 114: 169–183Google Scholar
  29. Franke WW (1974) Structure, biochemistry and functions of the nuclear envelope. Int. Rev. Cytol. 4: 72–236Google Scholar
  30. Garcia-Bustos JF, Wagner P & Hall MN (1991) Yeast cell-free nuclear protein import requires ATP hydrolysis. Exp. Cell Res. 192: 213–219Google Scholar
  31. Georgatos SD, Maroulakou I & Blobel G (1989) Lamin A, Lamin B, and Lamin B receptor analogues in yeast. J. Cell Biol. 108: 2069–2082Google Scholar
  32. Goldfarb D (1988) Karyophilic peptides: applications to the study of nuclear transport. Cell Biol. Inter. Reports. 12: 809–832Google Scholar
  33. Ghosh S & Baltimore D (1990) Activationin vitro of NF-ϰB by phosphorylation of its inhibitor IϰB. Nature 344: 678–682Google Scholar
  34. Greber UF, Senior A & Gerace L (1990) A major glycoprotein of the nuclear pore complex is a membrane-spanning polypeptide with a large luminal domain and a small cytoplasmic tail. EMBO J. 9: 1495–1502Google Scholar
  35. Hall MN, Hereford L & Herskowitz I (1984) Targeting ofE. coli b-galactosidase to the nucleus in yeast. Cell 36: 1057–1065Google Scholar
  36. Hall MN, Craik C & Hiraoka Y (1990) Homeodomain of yeast repressor alpha 2 contains a nuclear localization signal. Proc. Natl. Acad. Sci. USA 87: 6954–6958Google Scholar
  37. Hart GW, Haltiwanger RS, Holt GD & Kelly WG (1989) Glycosylation in the nucleus and cytoplasm. Ann. Rev. Biochem. 58: 841–874Google Scholar
  38. Hurt EC (1988) A novel nucleoskeletal-like protein located at the nuclear periphery is required for the life cycle ofSaccharomyces cerevisiae. EMBO J. 7: 4323–4334Google Scholar
  39. (1989) NSP1, a yeast protein located at the nuclear periphery, is required for the cell cycle ofSaccharomyces cerevisiae. J. Cell Sci. 12: 243–252Google Scholar
  40. (1990) Targeting of a cytosolic protein to the nuclear periphery. J. Cell Biol. 111: 2829–2837Google Scholar
  41. Hurt EC, McDowall A & Schimmang T (1988) Nucleolar and nuclear envelope proteins of the yeastSaccharomyces cerevisiae. Eur. J. Cell Biol. 46: 554–563Google Scholar
  42. Hurt EC, Mutvei A & Carmo-Fonseca M (1992) The nuclear envelope of the yeastS. cerevisiae. Int. Rev. Cytol. (in press)Google Scholar
  43. Jordan EG, Severs NJ & Williamson DH (1977) Nuclear pore formation and the cell cycle inSaccharomyces cerevisiae. Exp. Cell Res. 104: 446–449Google Scholar
  44. Kalderon D, Roberts BL, Richardson WP & Smith AE (1984) A short amino acid sequence able to specify nuclear location. Cell 39: 499–509Google Scholar
  45. Kalinich JF & Douglas MG (1989)In vitro translocation through the yeast nuclear envelope. J. Biol. Chem. 264: 17979–17989Google Scholar
  46. Lee WC & Melese T (1989) Identification and characterization of a nuclear localization sequence-binding protein in yeast. Proc. Natl. Acad. Sci. USA 86: 8808–8812Google Scholar
  47. Lee WC, Xue Z & Melese T (1991) TheNSR1 gene encodes a protein that specifically binds nuclear localization sequences and has two RNA recognition motifs. J. Cell Biol 113: 1–12Google Scholar
  48. Mann K & Mecke D (1980) Isolation and characterization of nuclei and nuclear membranes fromSaccharomyces cerevisiae protoplasts. FEBS Lett. 122: 95–99Google Scholar
  49. Mann K-H & Mecke D (1982) The isolation ofSaccharomyces cerevisiae nuclear membranes with nuclease and high-salt treatment. Biochim. Biophys. Acta 687: 57–62Google Scholar
  50. Maul GG (1977) The nuclear and cytoplasmic pore complex: structure, dynamics, distribution and evolution. Int. Rev. Cytol. 6: 75–186Google Scholar
  51. Meier UT & Blobel G (1990) A nuclear localization signal binding protein in the nucleolus. J. Cell Biol. 111: 2235–2245Google Scholar
  52. Moll T, Tebb G, Surana U, Robitsch H & Nasmyth K (1991) The role of phosphorylation and the CDC28 protein kinase in cell-cycle regulated nuclear import of theS. cerevisiae transcription factor SWI5. Cell 66: 743–758Google Scholar
  53. Moor H & Mühlethaler K (1963) Fine structure in frozen-etched yeast cells. J. Cell Biol. 17: 609–627Google Scholar
  54. Moreland RB, Nam HG, Hereford LM & Fried HM (1985) Identification of a nuclear localization signal of a yeast ribosomal protein. Proc. Natl. Acad. Sci. USA 82: 6561–6565Google Scholar
  55. Moreland RB, Langevin GL, Singer RH, Garcea RL & Hereford LM (1987) Amino acid sequences that determine the nuclear localization of yeast histone 2B. Mol. Cell Biol. 7: 4048–4057Google Scholar
  56. Nasmyth K, Seddon A & Ammerer G (1987) Cell cycle regulation of SW15 is required for mother-cell-specific HO transcription in yeast. Cell 49: 549–558Google Scholar
  57. Nasmyth K, Adolf G, Lydall D & Seddon A (1990) The identification of a second cell cycle control on the HO promoter in yeast: cell cycle regulation of SWI5 nuclear entry. Cell 62: 631–647Google Scholar
  58. Nehrbass U, Kern H, Mutvei A, Horstmann H, Marshallsay B & Hurt EC (1990) NSP1: A yeast nuclear envelope protein localized at the nuclear pores exerts its essential function by its carboxy-terminal domain. Cell 61: 979–989Google Scholar
  59. Nelson M & Silver P (1989) Context affects nuclear protein localization inSaccharomyces cerevisiae. Mol. Cell Biol. 9: 384–389Google Scholar
  60. Newmeyer DD & Forbes DJ (1988) Nuclear import can be separated into distinct stepsin vitro: nuclear pore binding and translocation. Cell 52: 641–653Google Scholar
  61. Park MK, D'Onofrio M, Willingham MC & Hanover JA (1987) A monoclonal antibody against a family of nuclear pore proteins (nucleoporins): O-linked N-acetylglucosamine is part of the immunodeterminant. Proc. Natl. Acad. Sci. USA 84: 6462–6466Google Scholar
  62. Peters R (1986) Fluorescence microphotolysis to measure nucleocytoplasmic transport and intracellular mobility. Biochim. Biophys. Acta 864: 305–359Google Scholar
  63. Picard D & Yamamoto KR (1987) Two signals mediate hormone-dependent nuclear localization of the glucocorticoid receptor. EMBO J. 6: 3333–3340Google Scholar
  64. Richardson WD, Mills AD, Dilworth SM, Laskey RA & Dingwall C (1988) Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by lower translocation through nuclear pores. Cell 52: 655–664Google Scholar
  65. Rihs H-P & Peters R (1989) Nuclear transport kinetics depend on phosphorylation site-containing sequences flanking the karyophilic signal of the SV40 T-antigen. EMBO J. 8: 1479–1484Google Scholar
  66. Rihs H-P, Jans DA, Fan H & Peters R (1991) The rate of nuclear cytoplasmic protein transport is determined by the casein kinase II site flanking the nuclear localization sequence of the SV40 T-antigen. EMBO J. 10: 633–639Google Scholar
  67. Robbins J, Dilworth SM, Laskey RA & Dingwall C (1991) Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: Identification of a class of bipartite nuclear targeting sequence. Cell 64: 615–623Google Scholar
  68. Roberts BL, Richardson WD & Smith AE (1987) The effect of protein context on nuclear location signal function. Cell 50: 465–475Google Scholar
  69. Rothblatt JA, Deshaies SL, Sanders SL, Daum G & Schekman R (1989) Multiple genes are required for proper insertion of secretory proteins into the endoplasmic reticulum in yeast. J. Cell Biol. 109: 2641–2652Google Scholar
  70. Sadler I, Chiang A, Kurihara T, Rothblatt J, Way J & Silver P (1989) A yeast gene important for protein assembly into the endoplasmic reticulum and the nucleus has homology to DnaJ, anEscherichia coli heat shock protein. J. Cell Biol. 109: 2665–2675Google Scholar
  71. Sanchez ER, Schlesinger MJ & Pratt WB (1985) The 90 kD non-steroid-binding phosphoprotein that binds to the untransformed glucocorticoid receptor in molybdate stabilized L-cell cytosol is the murine 90 kD heat shock protein. J. Biol. Chem. 260: 12398–12401Google Scholar
  72. Silver P, Sadler I & Osborne MA (1989) Yeast proteins that recognize nuclear localization sequences. J. Cell Biol. 109: 983–989Google Scholar
  73. Silver PA, Keegan LP & Ptashne M (1984) Amino terminus of the yeast GAL4 gene product is sufficient for nuclear localization. Proc. Natl. Acad. Sci. USA 81: 5951–5955Google Scholar
  74. Snow CM, Senior A & Gerace L (1987) Monoclonal antibodies identify a group of nuclear pore complex glycoproteins. J. Cell Biol. 104: 1143–1156Google Scholar
  75. Starr CM & Hanover JA (1991) A common structural motif in nuclear pore proteins (nucleoporins). BioEssays 13: 145–146Google Scholar
  76. Steinert PM & Roop DR (1988) Molecular and cellular biology of intermediate filaments. Ann. Rev. Biochem. 57: 593–625Google Scholar
  77. Steinert PM, Steven AC & Roop DR (1985) The molecular biology of intermediate filaments. Cell 42: 411–419Google Scholar
  78. Stochaj U, Osborne M, Kurihara T & Silver P (1991) A yeast protein that binds nuclear localization signals: Purification, localization, and antibody inhibition of binding activity. J. Cell Biol. 113: 1243–1254Google Scholar
  79. Underwood MR & Fried HM (1990) Characterization of nuclear localization sequences derived from the yeast ribosomal protein L29. EMBO J. 9: 91–99Google Scholar
  80. Wozniak RK, Bartnik E & Blobel G (1989) Primary structure analysis of an integral membrane glycoprotein of the nuclear pore. J. Cell Biol. 108: 2083–2092Google Scholar
  81. Yoneda Y, Imamoto-Sonobe N, Yamaizumi M & Uchida T (1987) Reversible inhibition of protein import into the nucleus by wheat germ agglutinin injected into cultured cells. Exp. Cell Res. 173: 586–595Google Scholar

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • U. Nehrbass
    • 1
  • E. C. Hurt
    • 1
  1. 1.European Molecular Biology LaboratoryHeidelbergGermany

Personalised recommendations