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Passive Transport of Macromolecules through Xenopus laevis Nuclear Envelope

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Abstract

Although nuclear pore complexes (NPC) are considered to be key structures in gene expression, little is known about their regulatory control. In order to explore the regulatory mechanism of passive transport of small macromolecules we examined the influence of different factors on the diffusional pathway of NPCs in isolated Xenopus laevis oocyte nuclei. Diffusion of fluorescence-labeled 10-kD dextran was measured across the nuclear envelope with confocal fluorescence microscopy. Surprisingly, the filling state of the perinuclear Ca2+ store had no influence on passive transport of 10-kD dextran. Furthermore, nuclear envelope permeability was independent of cytoplasmic pH (pH range 8.3–6.3). In contrast, nuclear swelling, induced by omission of the endogenous cytosolic macromolecules, clearly increased nuclear permeability. An antibody against the glycoprotein gp62, located at the central channel entrance, reduced macromolecule diffusion. In addition, nuclei from transcriptionally active, early developmental stages (stage II) were less permeable compared to transcriptionally inactive, late-developmental-stage (stage VI) nuclei. In stage II nuclei, atomic force microscopy disclosed NPC central channels with plugs that most likely were ribonucleoproteins exiting the nucleus. In conclusion, the difference between macromolecule permeability and previous measurements of electrical resistance strongly indicates separate routes for macromolecules and ions across the nuclear envelope.

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Acknowledgements

We thank Prof. G. Krohne, University of Würzburg, Germany, for the generous gift of gp62 antibody. We gratefully acknowledge the excellent technical assistance of Marianne Wilhelmi and Hannelore Arnold. The project was supported by grants of the Volkswagenstiftung (Project BD 151103) and the Interdisziplinäres Klinisches Forschungszentrum (IZKF Project A9).

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  1. Institute of Physiology I, Nanolab, University of Münster, D-48149 Münster, Germany

    K. Enss, T. Danker, A. Schlune, I. Buchholz & H. Oberleithner

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  1. K. Enss
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  2. T. Danker
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Correspondence to H. Oberleithner.

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Enss, K., Danker, T., Schlune, A. et al. Passive Transport of Macromolecules through Xenopus laevis Nuclear Envelope . J. Membrane Biol. 196, 147–155 (2003). https://doi.org/10.1007/s00232-003-0632-0

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  • DOI: https://doi.org/10.1007/s00232-003-0632-0

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