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The Hippo Pathway pp 115–119Cite as

Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1893))

Abstract

The translocation or shuttling of Hippo proteins between the nucleus and cytoplasm is a rapid event following cytoskeletal or mechanical cues as well as stimulation with extracellular growth factors. Here we describe an experimental procedure for a simple and fast separation of nuclear and cytoplasmic fractions which maintains protein integrity and integrity of protein-protein complexes, indicating that it should be applicable to many experimental questions.

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References

  1. Dupont S, Morsut L, Aragona M et al (2011) Role of YAP/TAZ in mechanotransduction. Nature 474:179–183. https://doi.org/10.1038/nature10137

    Article  CAS  PubMed  Google Scholar 

  2. Aragona M, Panciera T, Manfrin A et al (2013) A mechanical checkpoint controls multicellular growth through YAP/TAZ regulation by actin-processing factors. Cell 154:1047–1059. https://doi.org/10.1016/j.cell.2013.07.042

    Article  CAS  PubMed  Google Scholar 

  3. Wada K-I, Itoga K, Okano T et al (2011) Hippo pathway regulation by cell morphology and stress fibers. Development 138:3907–3914. https://doi.org/10.1242/dev.070987

    Article  CAS  PubMed  Google Scholar 

  4. Zhao B, Wei X, Li W et al (2007) Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control. Genes Dev 21:2747–2761. https://doi.org/10.1101/gad.1602907

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Varelas X, Samavarchi-Tehrani P, Narimatsu M et al (2010) The crumbs complex couples cell density sensing to Hippo-dependent control of the TGF-β-SMAD pathway. Dev Cell 19:831–844. https://doi.org/10.1016/j.devcel.2010.11.012

    Article  CAS  PubMed  Google Scholar 

  6. Thomasy SM, Morgan JT, Wood JA et al (2013) Substratum stiffness and latrunculin B modulate the gene expression of the mechanotransducers YAP and TAZ in human trabecular meshwork cells. Exp Eye Res 113:66–73. https://doi.org/10.1016/j.exer.2013.05.014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Graham JM, Rickwood D (1997) Subcellular fractionation : a practical approach. IRL Press at Oxford University Press, Oxford

    Google Scholar 

  8. Das A, Fischer RS, Pan D, Waterman CM (2016) YAP nuclear localization in the absence of cell-cell contact is mediated by a filamentous actin- dependent, myosin II-and phospho-YAP independent pathway during ECM mechanosensing. J Biol Chem 291(12):6096–6110. https://doi.org/10.1074/jbc.M115.708313

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Oncology, University of Oxford, Oxford, UK

    Maria Chatzifrangkeskou & Eric O’Neill

Authors
  1. Maria Chatzifrangkeskou
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  2. Eric O’Neill
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Corresponding author

Correspondence to Eric O’Neill .

Editor information

Editors and Affiliations

  1. Cancer Institute, University College London, London, UK

    Alexander Hergovich

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Chatzifrangkeskou, M., O’Neill, E. (2019). Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors. In: Hergovich, A. (eds) The Hippo Pathway. Methods in Molecular Biology, vol 1893. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8910-2_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8910-2_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8909-6

  • Online ISBN: 978-1-4939-8910-2

  • eBook Packages: Springer Protocols

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