Nuclear Pores: Methods for Preparation

Cécile Fradin, McMaster University, Hamilton, Ontario, Canada

Published online: July 2003

DOI: 10.1038/npg.els.0002706

Nuclear pore complexes are large protein assemblies that span the nuclear envelope and are involved in the regulation of molecular transport between the nucleus and the cytoplasm. There are three different methods that allow preparation of intact functional nuclear pore complexes: cell permeabilization, preparation of isolated nuclei, and cell-free nuclear reconstitution.

Keywords: nuclear pore complex; nuclear envelope; nucleus; nuclear transport; intracellular trafficking

Figure 1. A schematic representation of a nuclear pore complex embedded in the nuclear membrane. The cytoplasmic side of the membrane faces up; the nucleoplasmic side faces down. Courtesy of S. Beckermann.
Figure 2. Electron micrograph of a nuclear pore seen (a) from the nuclear side and (b) from the cytoplasmic side. Scale bar = 20 nm. Both micrographs are at the same scale. Reproduced from Allen TD, Cronshaw JM, Bagley S, Kiseleva E and Goldberg MW (2000) Journal of Cell Science 113: 1651–1659.
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 References
    Adam SA, Sterne-Marr R and Gerace L (1990) Nuclear protein import in permeabilized mammalian cells requires cytoplasmic factors. Journal of Cell Biology 111: 807–816.
    Goldberg MW and Allen TD (1992) High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores. Journal of Cell Biology 119: 1429–1440.
    Görlich D, Prehn S, Laskey RA and Hartmann E (1994) Isolation of a protein that is essential for the first step of nuclear protein import. Cell 79: 767–778.
    Keminer O and Peters R (1999) Permeability of single nuclear pores. Biophysical Journal 77: 217–228.
    Lohka MJ (1988) The reconstitution of nuclear envelopes in cell-free extracts. Cell Biology International Reports 12: 833–848.
    Newport (1987) Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. Cell 48: 205–217.
    Newport J and Spann T (1987) Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes. Cell 48: 219–230.
    Rout MP and Blobel G (1993) Isolation of the yeast nuclear pore complex. Journal of Cell Biology 123: 771–783.
 Further Reading
    Görlich D and Kutay U (1999) Transport between the cell nucleus and the cytoplasm. Annual Review of Cell and Developmental Biology 15: 607–660.
    Newmeyer DD and Wilson KL (1991) Egg extracts for nuclear import and nuclear assembly reactions. Methods in Cell Biology 36: 607–631.
    Rout MP, Aitchison JD, Suprapto A et al. (2000) The yeast nuclear pore complex: composition, architecture and transport mechanism. Journal of Cell Biology 148: 635–651.
    Strambio-de-Castillia C, Blobel G and Rout MP (1995) Isolation and characterization of nuclear envelopes from the yeast Saccharomyces. Journal of Cell Biology 131: 19–31.

Related Sub-Topics:

Cell Membranes | Cell Compartments and Cellular Organelles | Techniques in Cell Biology | Membrane Proteins
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Article Title: Nuclear Pores: Methods for Preparation
 
How to Cite close
Fradin, Cécile(Jul 2003) Nuclear Pores: Methods for Preparation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002706]