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Nuclear pore protein TPR associates with lamin B1 and affects nuclear lamina organization and nuclear pore distribution

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Abstract

The organization of the nuclear periphery is crucial for many nuclear functions. Nuclear lamins form dense network at the nuclear periphery and play a substantial role in chromatin organization, transcription regulation and in organization of nuclear pore complexes (NPCs). Here, we show that TPR, the protein located preferentially within the nuclear baskets of NPCs, associates with lamin B1. The depletion of TPR affects the organization of lamin B1 but not lamin A/C within the nuclear lamina as shown by stimulated emission depletion microscopy. Finally, reduction of TPR affects the distribution of NPCs within the nuclear envelope and the effect can be reversed by simultaneous knock-down of lamin A/C or the overexpression of lamin B1. Our work suggests a novel role for the TPR at the nuclear periphery: the TPR contributes to the organization of the nuclear lamina and in cooperation with lamins guards the interphase assembly of nuclear pore complexes.

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Acknowledgements

We thank to Iva Jelinkova for the help with immunofluorescence and HeLa cell culture. We thank the Microscopy Centre, Light Microscopy Core Facility, IMG ASCR, Prague, Czech Republic for the help with imaging, especially to Ivan Novotny for the help with microscope settings and acquisitions by SIM and STED.

Funding

This work was supported by the Grant Agency of the Czech Republic: [15-08835Y], [16-03346S], [16-03403S], [17-09103S], [18-19714S], by the institutional support of long-term conceptual support of development of the scientific organization (RVO: 68378050) and by the project “BIOCEV—Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” (CZ.1.05/1.1.00/02.0109), from the European Regional Development Fund. M.M. was supported by grant of ASCR (L200521801). J.U. was supported by GAUK (930218). We acknowledge the Light and Electron Microscopy Core Facility, IMG CAS supported by the MEYS CR (LM2015062), OPPK (CZ.2.16/3.1.00/21547), NPU I (LO1419) and ERDF (Project no. CZ.02.1.01/0.0/0.0/16_013/0001775).

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  1. Jindřiška Fišerová and Miloslava Maninová contributed equally to the work.

Authors and Affiliations

  1. Department of Biology of the Cell Nucleus, Institute of Molecular Genetics CAS, v.v.i., Vídeňská 1083, 142 00, Prague, Czech Republic

    Jindřiška Fišerová, Miloslava Maninová, Jana Uhlířová, Lenka Šebestová & Pavel Hozák

  2. Department of Cybernetics, Faculty of Electrical Engineering, Czech Technical University in Prague, Prague, Czech Republic

    Tomáš Sieger

  3. Microscopy Centre-LM and EM, Institute of Molecular Genetics CAS, v.v.i., Vídeňská 1083, 142 00, Prague, Czech Republic

    Michaela Efenberková, Martin Čapek & Pavel Hozák

  4. CLIP Laboratories, Department of Paediatric Haematology and Oncology, Second Faculty of Medicine, Charles University and University Hospital Motol Prague, V Úvalu 84, 150 06, Prague, Czech Republic

    Karel Fišer

  5. Division BIOCEV, Institute of Molecular Genetics CAS, v.v.i., Průmyslová 595, Vestec, 252 50, Prague, Czech Republic

    Pavel Hozák

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  1. Jindřiška Fišerová
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Contributions

JF was responsible for conception, JF and MM for design of the experiments, JF, MM, KF and PH for the biological data analysis; JF, MM, JU and LS for the experimental execution; ME for the Matlab programming, TS for statistical analysis; JF and MC for the image analysis, JF for drafting the article and JF, MM, TS, ME, MC, JU, LS, KF and PH for editing the article prior to submission.

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Correspondence to Jindřiška Fišerová.

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Fišerová, J., Maninová, M., Sieger, T. et al. Nuclear pore protein TPR associates with lamin B1 and affects nuclear lamina organization and nuclear pore distribution. Cell. Mol. Life Sci. 76, 2199–2216 (2019). https://doi.org/10.1007/s00018-019-03037-0

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