Reference - Detail
| RRC ID | 68794 |
|---|---|
| Author | Furuta M, Kose S, Kehlenbach RH, Imamoto N. |
| Title | Analysis of nucleocytoplasmic transport in digitonin-permeabilized cells under different cellular conditions. |
| Journal | Methods Cell Biol |
| Abstract |
The regulation of nucleocytoplasmic transport is crucial not only for basic cellular activities but also for physiological adaptation to specific situation during the cell cycle, development, or stress. Although a wide variety of transport pathways have been identified in eukaryotic cells, the functional significance of their multiplicity remains unclear. The best-characterized nuclear transport receptors (NTRs) are the members of the importin β family (karyopherin, transportin) whose association with specific cargoes is regulated by the GTPase Ran. In this chapter, we first provide an overview of the various expression vectors used to purify recombinant NTRs. We then describe two sets of recent examples of using well-established digitonin-permeabilized cell-free transport systems in mammalian cells to mimic different cellular conditions in living cells: normal/heat-shock conditions and interphase/mitosis. In the former case, physiological regulation impacts different transport pathways in opposite ways. In the latter case, the importin β-Ran system is used at different cell-cycle stages but with the same biochemical principle to specify the nuclear localization and chromatin loading of a specific protein, respectively. This in vitro transport assay, when adapted to specific cellular conditions or particular substrates, should help to uncover specific transport pathways or transport factors function under different cellular conditions. |
| Volume | 122 |
| Pages | 331-52 |
| Published | 2014-1-1 |
| DOI | 10.1016/B978-0-12-417160-2.00015-1 |
| PII | B978-0-12-417160-2.00015-1 |
| PMID | 24857737 |
| MeSH | Active Transport, Cell Nucleus / physiology* Cell Line, Tumor Cell-Free System DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism Digitonin / pharmacology* Escherichia coli / genetics HeLa Cells Humans Interphase / genetics Kinesins / genetics Kinesins / metabolism Mitosis / genetics Nuclear Pore / metabolism Signal Transduction alpha Karyopherins / metabolism beta Karyopherins / metabolism ran GTP-Binding Protein / metabolism* |
| IF | 1.441 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 2 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | HeLa-S3 |