Skip to main content
SpringerLink
Log in

Research Methodologies for the Investigation of Cell Nucleus

  • Chapter
Nuclear Import and Export in Plants and Animals

Part of the book series: Molecular Biology Intelligence Unit ((MBIU))

Abstract

It is impossible to detail all of the techniques used in the nuclear transport field within the allotted space. Since some methods (EM, cell and molecular biology, etc.) are dealt with in other Chapters, here I will focus on the approaches that are less familiar to the cell and molecular biologist (e.g., patch-clamp and atomic force microscopy). I believe that some of the observations I make are significant when interpreting experimental measurements.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Stauber RH. Methods and assays to investigate nuclear export. Curr Top Microbiol Immunol 2001; 259:119–128.

    PubMed  CAS  Google Scholar 

  2. PubMed: A service from the National Center for Biotechnology Information (NCBI), National Library of Medicine (NLM). National Institutes of Health (NIH), US government. NCBI Homepage

    Google Scholar 

  3. Fahrenkrog B, Stoffler D, Aebi U. Nuclear pore complex architecture and functional dynamics. Curr Top Microbiol Immunol 2001; 259:95–117.

    PubMed  CAS  Google Scholar 

  4. Stoffler D, Fahrenkrog B, Aebi U. The nuclear pore complex: from molecular architecture to functional dynamics. Curr Opin Cell Biol 1999; 11(3):391–401.

    Article  PubMed  CAS  Google Scholar 

  5. Allen TD, Cronshaw JM, Bagley S et al. The nuclear pore complex: mediator of translocation between nucleus and cytoplasm. J Cell Sci 2000; 113(Pt 10):1651–1659.

    PubMed  CAS  Google Scholar 

  6. Kiseleva E, Goldberg MW, Cronshaw J et al. The nuclear pore complex: structure, function, and dynamics. Crit Ver Eukaryot Gene Expr 2000; 10(1): 101–112.

    CAS  Google Scholar 

  7. Hitachi’s line of electron microscopes. Hitachi’s EM Line

    Google Scholar 

  8. Fruhwirth R, Regler M, Bock RK et al, eds. Data Analysis Techniques for High-Energy Physics. 2nd ed. Cambridge: Cambridge University Press, 2000:408.

    Google Scholar 

  9. Mazzanti M, Bustamante JO, Oberleithner H. Electrical dimension of the nuclear envelope. Physiol Ver 2001; 81(1):1–19.

    CAS  Google Scholar 

  10. Shahin V, Danker T, Enss K et al. Evidence for Ca2+-and ATP-sensitive peripheral channels in nuclear pore complexes. FASEB J 2001(11); 15:1895–1901.

    Article  PubMed  CAS  Google Scholar 

  11. Wang H, Clapham DE. Conformational changes of the in situ nuclear pore complex. Biophys J 1999; 77(1):241–247.

    PubMed  CAS  Google Scholar 

  12. Stoffler D, Goldie KN, Feja B et al. Calcium-mediated structural changes of native nuclear pore complexes monitored by time-lapse atomic force microscopy. J Mol Biol 1999; 287(4):741–752.

    Article  PubMed  CAS  Google Scholar 

  13. Hinshaw JE, Carragher BO, Milligan RA. Architecture and design of the nuclear pore complex. Cell 1992; 69(7):1133–1141.

    Article  PubMed  CAS  Google Scholar 

  14. Bustamante JO, Liepins A, Prendergast RA et al. Patch clamp and atomic force microscopy demonstrate TATA-binding protein (TBP) interactions with the nuclear pore complex. J Membr Biol 1995; 146:263–272.

    PubMed  CAS  Google Scholar 

  15. Monnè L. Permeability of the nuclear membrane to vital stains. Proc Soc Exptl Biol Med 1935; 32:1197–1199.

    Google Scholar 

  16. Molecular Probes’ line of fluorescent probes. Molecular Probes Homepage

    Google Scholar 

  17. Bustamante JO, Michelette ER, Geibel JP et al. Dendrimer-assisted patch-damp sizing of nuclear pores. Pflügers Arch 2000; 439(6):829–837.

    Article  PubMed  CAS  Google Scholar 

  18. Bustamante JO, Hanover JA, Liepins A. The ion channel behavior of the nuclear pore complex. J Membr Biol 1995; 146:239–251.

    PubMed  CAS  Google Scholar 

  19. Clontech’s lines of products related to GFP. Clontech’s Homepage

    Google Scholar 

  20. Ayoob JC, Sanger JM, Sanger JW. Visualization of the expression of green fluorescent protein (GFP)-linked proteins. Methods Mol Biol 2000; 137:153–157.

    PubMed  CAS  Google Scholar 

  21. Tomura A, Goto K, Morinaga H et al. The subnuclear three-dimensional image analysis of androgen receptor fused to green fluorescence protein. J Biol Chem 2001; 276:28395–28401.

    Article  PubMed  CAS  Google Scholar 

  22. Terskikh A, Fradkov A, Ermakova G et al. “Fluorescent timer”: Protein that changes color with time. Science 2000; 290(5496):1585–1588.

    Article  PubMed  CAS  Google Scholar 

  23. Stage-Zimmermann T, Schmidt U, Silver PA. Factors affecting nuclear export of the 60S ribosomal subunit in vivo. Mol Biol Cell 2000; 11:3777–3789.

    PubMed  CAS  Google Scholar 

  24. Perez-Terzic C, Stehno-Bittel L, Clapham DE. Nucleoplasmic and cytoplasmic differences in the fluorescence properties of the calcium indicator Fluo-3. Cell Calcium 1997; 21:275–282.

    Article  PubMed  CAS  Google Scholar 

  25. Badminton MN, Kendall JM, Rembold CM et al. Current evidence suggests independent regulation of nuclear calcium. Cell Calcium 1998; 23:79–86.

    Article  PubMed  CAS  Google Scholar 

  26. Keminer O, Peters R. Permeability of single nuclear pores. Biophys J 1999; 77:217–228.

    PubMed  CAS  Google Scholar 

  27. Keminer O, Siebrasse JP, Zerf K et al. Optical recording of signal-mediated protein transport through single nuclear pore complexes. Proc Natl Acad Sci USA 1999; 96:11842–11847.

    Article  PubMed  CAS  Google Scholar 

  28. Kues T, Peters R, Kubitscheck U. Visualization and tracking of single protein molecules in the cell nucleus. Biophys J 2001; 80:2954–2967.

    PubMed  CAS  Google Scholar 

  29. Radtke T, Schmalz D, Coutavas E et al. Kinetics of protein import into isolated Xenopus oocyte nuclei. Proc Natl Acad Sci USA 2001; 98:2407–2412.

    Article  PubMed  CAS  Google Scholar 

  30. Cullen BR. Journey to the center of the cell. Cell 2001; 105:697–700.

    Article  PubMed  CAS  Google Scholar 

  31. Liu J, Xiao N, DeFranco DB. Use of digitonin-permeabilized cells in studies of steroid receptor subnuclear trafficking. Methods 1999; 19:403–409.

    Article  PubMed  CAS  Google Scholar 

  32. Guan T, Kehlenbach RH, Schirmer EC et al. Nup50, a nucleoplasmically oriented nucleoporin with a role in nuclear protein export. Mol Cell Biol 2000; 20:5619–5630.

    Article  PubMed  CAS  Google Scholar 

  33. Petersen JM, Her LS, Dahlberg JE. Multiple vesiculoviral matrix proteins inhibit both nuclear export and import. Proc Natl Acad Sci USA 2001; 98:8590–8595.

    Article  PubMed  CAS  Google Scholar 

  34. Patterson SD. Mass spectrometry and proteomics. Physiol Genomics 2000; 2(2):59–65.

    PubMed  CAS  Google Scholar 

  35. Rappsilber J, Siniossoglou S, Hurt EC et al. A generic strategy to analyze the spatial organization of multi-protein complexes by cross-linking and mass spectrometry. Anal Chem 2000; 72:267–275.

    Article  PubMed  CAS  Google Scholar 

  36. Rout MP, Aitchison JD, Suprapto A et al. The yeast nuclear pore complex: Composition, architecture, and transport mechanism. J Cell Biol 2000; 148(4):635–651.

    Article  PubMed  CAS  Google Scholar 

  37. Funakoshi M, Tsuchiya Y, Arahata K. Emerin and cardiomyopathy in Emery-Dreifuss muscular dystrophy. Neuromuscul Disord 1999; 9(2):108–114.

    Article  PubMed  CAS  Google Scholar 

  38. Haraguchi T, Koujin T, Hayakawa T et al. Live fluorescence imaging reveals early recruitment of emerin, LBR, RanBP2, and Nup153 to reforming functional nuclear envelopes. J Cell Sci 2000; 113 (Pt 5):779–794.

    PubMed  CAS  Google Scholar 

  39. Ahuja HG, Popplewell L, Tcheurekdjian L et al. NUP98 gene rearrangements and the clonal evolution of chronic myelogenous leukemia. Genes Chromosomes Cancer 2001; 30(4):410–415.

    Article  PubMed  CAS  Google Scholar 

  40. Burke B, Mounkes LC, Stewart CL. The nuclear envelope in muscular dystrophy and cardiovascular diseases. Traffic 2001; 2(10):675–683.

    Article  PubMed  CAS  Google Scholar 

  41. Hegele RA, Yuen J, Cao H. Single-nucleotide polymorphisms of the nuclear lamina proteome. J Hum Genet 2001; 46:351–354.

    Article  PubMed  CAS  Google Scholar 

  42. Invernizzi P, Podda M, Battezzati PM et al. Autoantibodies against nuclear pore complexes are associated with more active and severe liver disease in primary biliary cirrhosis. J Hepatol 2001; 34(3):366–372.

    Article  PubMed  CAS  Google Scholar 

  43. Jaju RJ, Fidler C, Haas OA et al. A novel gene, NSD1, is fused to NUP98 in the t(5;11)(q35;p15.5) in de novo childhood acute myeloid leukemia. Blood 2001; 98(4): 1264–1267.

    Article  PubMed  CAS  Google Scholar 

  44. Nesher G, Margalit R, Ashkenazi YJ. Anti-nuclear envelope antibodies: Clinical associations. Semin Arthritis Rheum 2001; 30(5):313–320.

    Article  PubMed  CAS  Google Scholar 

  45. Perez-Terzic C, Gacy AM, Bortolon R et al. Directed inhibition of nuclear import in cellular hypertrophy. J Biol Chem 2001; 276(23):20566–20571.

    Article  PubMed  CAS  Google Scholar 

  46. Lu D, Yang H, Shaw G, Raizada MK. Angiotensin II-induced nuclear targeting of the angiotensin type 1 (AT1) receptor in brain neurons. Endocrinology 1998; 139(1):365–375.

    Article  PubMed  CAS  Google Scholar 

  47. Bhattacharya M, Peri KG, Almazan G et al. Nuclear localization of prostaglandin E2 receptors. Proc Natl Acad Sci USA 1998; 95(26):15792–157927.

    Article  PubMed  CAS  Google Scholar 

  48. Bustamante JO. Nuclear electrophysiology. J Membr Biol 1994; 138(2):105–112.

    PubMed  CAS  Google Scholar 

  49. Bustamante JO, Liepins A, Hanover JA. Nuclear pore complex ion channels. Mol Membr Biol 1994; 11(3):141–150.

    Article  PubMed  CAS  Google Scholar 

  50. The Nobel Prize in Physiology or Medicine 1963: Nobel Lectures of Hodgkin and Huxley. Nobel Prize for 1963

    Google Scholar 

  51. Bustamante JO. Restricted ion flow at the nuclear envelope of cardiac myocytes. Biophys J 1993; 64(6):1735–1749.

    Article  PubMed  CAS  Google Scholar 

  52. Stehno-Bittel L, Luckhoff A, Clapham DE. Calcium release from the nucleus by InsP3 receptor channels. Neuron 1995; 14(1):163–167.

    Article  PubMed  CAS  Google Scholar 

  53. Mak DO, Foskett JK. Single-channel kinetics, inactivation, and spatial distribution of inositol trisphosphate IP3 receptors in Xenopus oocyte nucleus. J Gen Physiol 1997; 109(5):571–587.

    Article  PubMed  CAS  Google Scholar 

  54. Mak DO, McBride S, Foskett JK. ATP regulation of type 1 inositol 1,4,5-trisphosphate receptor channel gating by allosteric tuning of Ca2+ activation. J Biol Chem 1999; 274(32):22231–22237.

    Article  PubMed  CAS  Google Scholar 

  55. Humbert JP, Matter N, Artault JC et al. Inositol 1,4,5-trisphosphate receptor is located to the inner nuclear membrane vindicating regulation of nuclear calcium signaling by inositol 1,4,5-trisphosphate. Discrete distribution of inositol phosphate receptors to inner and outer nuclear membranes. J Biol Chem 1996; 271(1):478–485.

    Article  PubMed  CAS  Google Scholar 

  56. Khoo KM, Han MK, Park JB et al. Localization of the cyclic ADP-ribose-dependent calcium signaling pathway in hepatocyte nucleus. J Biol Chem 2000; 275(32):24807–24817.

    Article  PubMed  CAS  Google Scholar 

  57. Strambio-de-Castillia C, Blobel G, Rout MP. Isolation and characterization of nuclear envelopes from the yeast Saccharomyces. J Cell Biol 1995; 131(1):19–31.

    Article  PubMed  CAS  Google Scholar 

  58. Lacinova L, Cleemann L, Morad M. Ca2+ channel modulating effects of heparin in mammalian cardiac myocytes. J Physiol 1993; 465:181–201.

    PubMed  CAS  Google Scholar 

  59. Tabares L, Mazzanti M, Clapham DE. Chloride channels in the nuclear membrane. J Membr Biol 1991; 123(1):49–54.

    Article  PubMed  CAS  Google Scholar 

  60. Franco-Obregon A, Wang HW, Clapham DE. Distinct ion channel classes are expressed on the outer nuclear envelope of T-and B-lymphocyte cell lines. Biophys J 2000; 79(1):202–214.

    PubMed  CAS  Google Scholar 

  61. Bustamante JO. Nuclear ion channels in cardiac myocytes. Pflügers Arch 1992; 421(5):473–485.

    Article  PubMed  CAS  Google Scholar 

  62. Loewenstein WR, Kanno Y, Ito S. Permeability of nuclear membranes. Ann NY Acad Sci 1966; 137(2):708–716.

    Article  PubMed  CAS  Google Scholar 

  63. Overbeek JThG. The Donnan equilibrium. Progr Biophys Biophys Chem 1956; 6:57–84.

    CAS  Google Scholar 

  64. Bustamante JO, McDonald TF. Sodium currents in segments of human heart cells. Science 1983; 220(4594):320–321.

    Article  PubMed  CAS  Google Scholar 

  65. Bustamante JO, Varanda WA. Patch-clamp detection of macromolecular translocation along nuclear pores. Braz J Med Biol Res 1998; 31(3):333–354.

    Article  PubMed  CAS  Google Scholar 

  66. Bustamante JO, Oberleithner H, Hanover JA et al. Patch clamp detection of transcription factor translocation along the nuclear pore complex channel. J Membr Biol 1995; 146(3):253–261.

    PubMed  CAS  Google Scholar 

  67. Bustamante JO, Michelette ER, Geibel JP et al. Calcium, ATP and nuclear pore channel gating. Pflügers Arch 2000; 439(4):433–444.

    Article  PubMed  CAS  Google Scholar 

  68. Danker T, Schillers H, Storck J et al. Nuclear hourglass technique: An approach that detects electrically open nuclear pores in Xenopus laevis oocyte. Proc Natl Acad Sci USA 1999; 96(23):13530–13535.

    Article  PubMed  CAS  Google Scholar 

  69. Kostyuk PC Intracellular perfusion. Annu Rev Neurosci 1982; 5:107–120.

    Article  PubMed  CAS  Google Scholar 

  70. The Nobel Prize in Physiology or Medicine 1991: Nobel Lectures of Neher and Sakmann. Nobel Prize for 1991

    Google Scholar 

  71. Feldherr CM, Akin D. The location of the transport gate in the nuclear pore complex. J Cell Sci 1997; 110(Pt 24):3065–3070.

    PubMed  CAS  Google Scholar 

  72. Celera’s animated primer on genomics. Celera’s Educational Link

    Google Scholar 

  73. Allen NP, Huang L, Burlingame A et al. Proteomic analysis of nucleoporin interacting proteins. J Biol Chem 2001; 276(31):29268–29274.

    Article  PubMed  CAS  Google Scholar 

  74. Adam SA. The nuclear pore complex. Genome Biol 2001; 2:REVIEWS0007.

    Google Scholar 

  75. Lewis TS, Hunt JB, Aveline LD et al. Identification of novel MAP kinase pathway signaling tar gets by functional proteomics and mass spectrometry. Mol Cell 2000; 6:1343–1354.

    Article  PubMed  CAS  Google Scholar 

  76. Bayliss R, Kent HM, Corbett AH et al. Crystallization and initial X-ray diffraction characterization of complexes of FxFG nucleoporin repeats with nuclear transport factors. J Struct Biol 2000; 131(3):240–247.

    Article  PubMed  CAS  Google Scholar 

  77. Kasianowicz JJ, Brandin E, Branton D et al. Characterization of individual polynucleotide molecules using a membrane channel. Proc Natl Acad Sci USA 1996; 93(24): 13770–13773.

    Article  PubMed  CAS  Google Scholar 

  78. Deamer DW, Akeson M. Nanopores and nucleic acids: prospects for ultrarapid sequencing. Trends Biotechnol 2000; 18(4):147–151.

    Article  PubMed  CAS  Google Scholar 

  79. Wilmut I, Schnieke AE, McWhir J et al. Viable offspring derived from fetal and adult mammalian cells. Nature 1997; 385(6619):810–813.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Nuclear Physiology Lab and Nanobiotechnology Group, Department of Physics, Univeridade Federal de Sergipe, The Brazilian Millenium Institute of Nanosciences, The Brazilian Nanosciences & Nanotechnology Network, Sao Cristovac, Brazil

    Jose Omar Bustamante

Authors
  1. Jose Omar Bustamante
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Eurekah.com and Kluwer Academic / Plenum Publishers

About this chapter

Cite this chapter

Bustamante, J.O. (2005). Research Methodologies for the Investigation of Cell Nucleus. In: Nuclear Import and Export in Plants and Animals. Molecular Biology Intelligence Unit. Springer, Boston, MA. https://doi.org/10.1007/0-387-27747-1_13

Download citation

Publish with us

Policies and ethics

Search

Navigation