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Analysis of nuclear actin by overexpression of wild-type and actin mutant proteins

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Abstract

Compared to the cytoplasmic F-actin abundance in cells, nuclear F-actin levels are generally quite low. However, nuclear actin is present in certain cell types including oocytes and under certain cellular conditions including stress or serum stimulation. Currently, the architecture and polymerization status of nuclear actin networks has not been analyzed in great detail. In this study, we investigated the architecture and functions of such nuclear actin networks. We generated nuclear actin polymers by overexpression of actin proteins fused to a nuclear localization signal (NLS). Raising nuclear abundance of a NLS wild-type actin, we observed phalloidin- and LifeAct-positive actin bundles forming a nuclear cytoskeletal network consisting of curved F-actin. In contrast, a polymer-stabilizing actin mutant (NLS-G15S-actin) deficient in interacting with the actin-binding protein cofilin generated a nuclear actin network reminiscent of straight stress fiber-like microfilaments in the cytoplasm. We provide a first electron microscopic description of such nuclear actin polymers suggesting bundling of actin filaments. Employing different cell types from various species including neurons, we show that the morphology of and potential to generate nuclear actin are conserved. Finally, we demonstrate that nuclear actin affects cell function including morphology, serum response factor-mediated gene expression, and herpes simplex virus infection. Our data suggest that actin is able to form filamentous structures inside the nucleus, which share architectural and functional similarities with the cytoplasmic F-actin.

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Acknowledgments

B.K. is supported by the DFG (Deutsche Forschungsgemeinschaft), Schram, Gottschalk, and Gemeinnützige Hertie foundation. G.P. and J.W. are supported by the DFG (priority program 1464). The authors declare no conflict of interest.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Institute for Physiological Chemistry, Ulm University, Albert-Einstein-Allee 11, 89081, Ulm, Germany

    Enikö Kokai, Henning Beck, Daniela Sinske, Ulrike Sebert & Bernd Knöll

  2. Institute for Physiological Chemistry, Martin Luther University Halle-Wittenberg, Hollystr. 1, 06097, Halle (Saale), Germany

    Julia Weissbach & Guido Posern

  3. Institute of Molecular Virology, Ulm University, Albert-Einstein-Allee 11, 89081, Ulm, Germany

    Franziska Arnold & Jan Münch

  4. Institute of Stochastics, Ulm University, Helmholtzstr. 18, 89069, Ulm, Germany

    Gerd Gaiselmann & Volker Schmidt

  5. Central Facility for Electron Microscopy, Ulm University, Albert-Einstein-Allee 11, 89081, Ulm, Germany

    Paul Walther

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  1. Enikö Kokai
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  2. Henning Beck
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  11. Guido Posern
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Correspondence to Bernd Knöll.

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Kokai, E., Beck, H., Weissbach, J. et al. Analysis of nuclear actin by overexpression of wild-type and actin mutant proteins. Histochem Cell Biol 141, 123–135 (2014). https://doi.org/10.1007/s00418-013-1151-4

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