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Differential basal-to-apical accessibility of lamin A/C epitopes in the nuclear lamina regulated by changes in cytoskeletal tension

Nature Materials volume 14, pages 1252–1261 (2015)Cite this article

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Abstract

Nuclear lamins play central roles at the intersection between cytoplasmic signalling and nuclear events. Here, we show that at least two N- and C-terminal lamin epitopes are not accessible at the basal side of the nuclear envelope under environmental conditions known to upregulate cell contractility. The conformational epitope on the Ig-domain of A-type lamins is more buried in the basal than apical nuclear envelope of human mesenchymal stem cells undergoing osteogenesis (but not adipogenesis), and in fibroblasts adhering to rigid (but not soft) polyacrylamide hydrogels. This structural polarization of the lamina is promoted by compressive forces, emerges during cell spreading, and requires lamin A/C multimerization, intact nucleoskeleton–cytoskeleton linkages (LINC), and apical-actin stress-fibre assembly. Notably, the identified Ig-epitope overlaps with emerin, DNA and histone binding sites, and comprises various laminopathy mutation sites. Our findings should help decipher how the physical properties of cellular microenvironments regulate nuclear events.

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Figure 1: Some, but not all, lamin A/C antibody immunostains show a basal-to-apical polarization of the nuclear envelope (NE).
Figure 2: Multimerization-deficient lamin A mutant (ΔN20-terminal deletion) strongly reduces the basal-to-apical polarization of the LA/C-C immuno stain.
Figure 3: Cell spreading coincides with the build-up of the basal-to-apical polarization of the LA/C-C and LA/C-N immunostains.
Figure 4: Polarization of the LA/C-C stain can be altered by tuning engineered two-dimensional cell environments and requires physically intact LINC-complexes.
Figure 5: Differential LA/C-C epitope accessibility is not altered when the cell nucleus is bent over a micropost, but can be re-established by cell compression using a non-adhesive PAA cushion.
Figure 6: Basal-to-apical polarization of the LA/C-C stains is reduced for pre-adipogenic hMSCs.

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Acknowledgements

We thank M. Burkhardt, J. L. B. Khuan (IMRE, Singapore) and S. Früh for their valuable help with micropost and adhesive microisland substrates; M. Vihinen-Ranta and D. Hodzic for EGFP-lamin A and EGFP-KASH2 constructs, respectively; R. Foisner, O. Medalia, J. Lammerding and R. Fässler for LMNA null, VIM null, EMD null and WT MEF cells; U. Kutay for SUN2 antibody; K. Maniura for the usage of the Amaxa Nucleofector II system. This research was supported by Foundations’ Post Doc Pool, Finland (T.O.I.); Academy of Finland, grants 252225 and 267471 (T.O.I.); Portuguese Foundation for Science and Technology, doctoral grant SFRH/BD/42019/2007 (L.A.); SystemsX.ch - ‘PhosphoNetX’, Internal Nr. 2-67124-08 (V.V.); ERC Advanced Grant European Community, GA233157 (V.V.); and SNF Swiss National Science Foundation, Grant 310030B_133122 (V.V.). Computational resources provided by the Swiss National Supercomputing Center (CSCS) and the University of Tampere Imaging Facility are gratefully acknowledged.

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  1. Teemu O. Ihalainen and Lina Aires: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Health Sciences and Technology, Laboratory of Applied Mechanobiology, ETH Zurich, 8093 Zurich, Switzerland

    Teemu O. Ihalainen, Lina Aires, Florian A. Herzog, Ruth Schwartlander, Jens Moeller & Viola Vogel

  2. NeuroGroup, BioMediTech, University of Tampere, 33014 Tampere, Finland

    Teemu O. Ihalainen

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Contributions

T.O.I., L.A. and V.V. designed the experiments. T.O.I. performed the PAA cushion experiments and L.A. carried out the LA/C-C epitope mapping. T.O.I. and L.A. did the remaining experiments with the help of J.M. (μ-island experiments) and R.S. (cell spreading experiments). F.H. was responsible for the structural analysis of lamin A and SMD simulations. All of the authors were involved in the analyses and interpretation of data. T.O.I., L.A. and V.V. wrote the paper, with the help of the co-authors.

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Correspondence to Viola Vogel.

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Ihalainen, T., Aires, L., Herzog, F. et al. Differential basal-to-apical accessibility of lamin A/C epitopes in the nuclear lamina regulated by changes in cytoskeletal tension. Nature Mater 14, 1252–1261 (2015). https://doi.org/10.1038/nmat4389

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