Abstract
The genome in eukaryotic cells is encased by two intricate and interconnected concentric membranes, which together with the underlying nuclear lamina form the nuclear envelope (NE). Two fundamental macromolecular structures are embedded within the nuclear envelope: the nuclear pore (NPC) and the LINC complex. The former perforates the nucleus controlling biomolecule trafficking between the nucleoplasm and the cytoplasm, while the latter integrates the nucleus via the cytoskeleton to the extracellular matrix. LINC complex structural and functional integrity is of utmost importance for various fundamental cellular functions. Mechanical forces are relayed into the nuclear interior via the LINC complex, which controls lamina organization, chromosome dynamics, and genome organization and stability. Thus, LINC constituents play pivotal roles in cellular architecture including organelle positioning, cell movement, tissue assembly, organ homeostasis, and organismal aging. The LINC complex oligomeric core contains several multi-isomeric, multifunctional, and often tissue-specific proteins. Therefore, for a proper functional analysis, genetic mouse models are an invaluable resource. Herein, we focus on the LINC complex roles in the skin and describe methods that enable the successful isolation of primary embryonic fibroblast and newborn skin cells, which can be then investigated functionally in vitro.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Crisp M, Liu Q, Roux K et al (2006) Coupling of the nucleus and cytoplasm: role of the LINC complex. J Cell Biol 172:41–53
Xiong H, Rivero F, Euteneuer U et al (2008) Dictyostelium Sun-1 connects the centrosome to chromatin and ensures genome stability. Traffic 9:708–724
Haque F, Mazzeo D, Patel JT et al (2010) Mammalian SUN protein interaction networks at the inner nuclear membrane and their role in laminopathy disease processes. J Biol Chem 285:3487–3498
Li P, Noegel AA (2015) Inner nuclear envelope protein SUN1 plays a prominent role in mammalian mRNA export. Nucleic Acids Res 43:9874–9888
Luxton GW, Starr DA (2014) KASHing up with the nucleus: novel functional roles of KASH proteins at the cytoplasmic surface of the nucleus. Curr Opin Cell Biol 28:69–75
Worman HJ (2012) Nuclear lamins and laminopathies. J Pathol 226:316–325
Sosa BA, Kutay U, Schwartz TU (2013) Structural insights into LINC complexes. Curr Opin Struct Biol 23:285–291
Rothballer A, Kutay U (2013) The diverse functional LINCs of the nuclear envelope to the cytoskeleton and chromatin. Chromosoma 122:415–429
Lattanzi G, Marmiroli S, Facchini A et al (2012) Nuclear damages and oxidative stress: new perspectives for laminopathies. Eur J Histochem e45:56
Rajgor D, Mellad JA, Autore F et al (2012) Multiple novel nesprin-1 and nesprin-2 variants act as versatile tissue-specific intracellular scaffolds. PLoS One 7:e40098
Zhen YY, Libotte T, Munck M et al (2002) NUANCE, a giant protein connecting the nucleus and actin cytoskeleton. J Cell Sci 115:3207–3222
Padmakumar VC, Abraham S, Braune S et al (2004) Enaptin, a giant actin-binding protein, is an element of the nuclear membrane and the actin cytoskeleton. Exp Cell Res 295:330–339
Wilhelmsen K, Ketema M, Truong H et al (2006) KASH-domain proteins in nuclear migration, anchorage and other processes. J Cell Sci 119:5021–5029
Lüke Y, Zaim H, Karakesisoglou I et al (2007) Nesprin-2 Giant (NUANCE) maintains nuclear envelope architecture and composition in skin. J Cell Sci 121:1887–1898
Rashmi RN, Eckes B, Glöckner G et al (2012) The nuclear envelope protein Nesprin-2 has roles in cell proliferation and differentiation during wound healing. Nucleus 3:172–186
Bhattacharya K, Swaminathan K, Peche VS et al (2016) Novel Coronin7 interactions with Cdc42 and N-WASP regulate actin organization and Golgi morphology. Sci Rep 6:25411
Zhang ZG, Bothe I, Hirche F et al (2006) Interactions of primary fibroblasts and keratinocytes with extracellular matrix proteins: contribution of alpha2beta1 integrin. J Cell Sci 119:1886–1895
Carthew J, Karakesisoglou I (2016) Detection of diverse and high molecular weight Nesprin-1 and Nesprin-2 isoforms using Western blotting. Methods Mol Biol 1411:221–232
Acknowledgments
This work was supported by Cologne Cluster of Excellence on Cellular Stress Responses in Aging-Associated Diseases (CECAD), the International Graduate School in Development Health and Disease (IGSDHD), and the BBSRC (Biotechnology and Biological Sciences Research Council).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Karakesisoglou, I., Mroß, C., Noegel, A.A. (2018). Functional Analysis of LINC Complexes in the Skin. In: Gundersen, G., Worman, H. (eds) The LINC Complex. Methods in Molecular Biology, vol 1840. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8691-0_20
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8691-0_20
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8690-3
Online ISBN: 978-1-4939-8691-0
eBook Packages: Springer Protocols