Measuring the Kinase Activities of the LATS/NDR Protein Kinases

  • Alexander HergovichEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1893)


The Hippo tissue growth control and regeneration pathway is a main regulator of the YAP/TAZ effectors. In this regard, the LATS/NDR serine/threonine protein kinases can function as central components of the Hippo core module. More specifically, LATS/NDR-mediated phosphorylation of YAP/TAZ on different residues can regulate the subcellular localization and/or stability of YAP/TAZ. Therefore, the assessment of LATS/NDR activities can serve as readout for the activity status of the Hippo pathway. Here, we describe our preferred methodology regarding the measurement of the activities of LATS/NDR kinases.

Key words

Hippo pathway Protein kinases Kinase assays Kinase substrates LATS1 LATS2 NDR1 NDR2 STK38 STK38L 



The Hergovich laboratory was supported by the Wellcome Trust (090090/Z/09/Z), BBSRC (BB/I021248/1), Worldwide Cancer Research (AICR; 11-0634), UCL Cancer Research UK Centre, and the National Institute for Health Research University College London Hospitals Biomedical Research Centre.


  1. 1.
    Harvey KF, Zhang X, Thomas DM (2013) The Hippo pathway and human cancer. Nat Rev Cancer 13(4):246–257. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Johnson R, Halder G (2014) The two faces of Hippo: targeting the Hippo pathway for regenerative medicine and cancer treatment. Nat Rev Drug Discov 13(1):63–79. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Meng Z, Moroishi T, Guan KL (2016) Mechanisms of Hippo pathway regulation. Genes Dev 30(1):1–17. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Yu FX, Zhao B, Guan KL (2015) Hippo pathway in organ size control, tissue homeostasis, and cancer. Cell 163(4):811–828. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Irvine KD, Harvey KF (2015) Control of organ growth by patterning and hippo signaling in Drosophila. Cold Spring Harb Perspect Biol 7(6):a019224CrossRefGoogle Scholar
  6. 6.
    Pan D (2010) The hippo signaling pathway in development and cancer. Dev Cell 19(4):491–505CrossRefGoogle Scholar
  7. 7.
    Hergovich A, Stegert MR, Schmitz D, Hemmings BA (2006) NDR kinases regulate essential cell processes from yeast to humans. Nat Rev Mol Cell Biol 7(4):253–264CrossRefGoogle Scholar
  8. 8.
    Pearce LR, Komander D, Alessi DR (2010) The nuts and bolts of AGC protein kinases. Nat Rev Mol Cell Biol 11(1):9–22CrossRefGoogle Scholar
  9. 9.
    Avruch J, Zhou D, Fitamant J, Bardeesy N, Mou F, Barrufet LR (2012) Protein kinases of the Hippo pathway: regulation and substrates. Semin Cell Dev Biol 23(7):770–784CrossRefGoogle Scholar
  10. 10.
    Hergovich A (2011) MOB control: reviewing a conserved family of kinase regulators. Cell Signal 23(9):1433–1440. CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Sudol M, Harvey KF (2010) Modularity in the Hippo signaling pathway. Trends Biochem Sci 35(11):627–633CrossRefGoogle Scholar
  12. 12.
    Manning G, Whyte DB, Martinez R, Hunter T, Sudarsanam S (2002) The protein kinase complement of the human genome. Science 298(5600):1912–1934CrossRefGoogle Scholar
  13. 13.
    Hergovich A (2016) The roles of ndr protein kinases in hippo signalling. Genes 7(5):21CrossRefGoogle Scholar
  14. 14.
    Dong J, Feldmann G, Huang J, Wu S, Zhang N, Comerford SA, Gayyed MF, Anders RA, Maitra A, Pan D (2007) Elucidation of a universal size-control mechanism in Drosophila and mammals. Cell 130(6):1120–1133CrossRefGoogle Scholar
  15. 15.
    Hao Y, Chun A, Cheung K, Rashidi B, Yang X (2008) Tumor suppressor LATS1 is a negative regulator of oncogene YAP. J Biol Chem 283(9):5496–5509CrossRefGoogle Scholar
  16. 16.
    Lei Q-Y, Zhang H, Zhao B, Zha Z-Y, Bai F, Pei X-H, Zhao S, Xiong Y, Guan K-L (2008) TAZ promotes cell proliferation and epithelial-mesenchymal transition and is inhibited by the hippo pathway. Mol Cell Biol 28(7):2426–2436CrossRefGoogle Scholar
  17. 17.
    Liu C-Y, Zha Z-Y, Zhou X, Zhang H, Huang W, Zhao D, Li T, Chan SW, Lim CJ, Hong W (2010) The hippo tumor pathway promotes TAZ degradation by phosphorylating a phosphodegron and recruiting the SCFβ-TrCP E3 ligase. J Biol Chem 285(48):37159–37169CrossRefGoogle Scholar
  18. 18.
    Zhao B, Li L, Tumaneng K, Wang C-Y, Guan K-L (2010) A coordinated phosphorylation by Lats and CK1 regulates YAP stability through SCFβ-TRCP. Genes Dev 24(1):72–85CrossRefGoogle Scholar
  19. 19.
    Zhao B, Wei X, Li W, Udan RS, Yang Q, Kim J, Xie J, Ikenoue T, Yu J, Li L (2007) Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control. Genes Dev 21(21):2747–2761CrossRefGoogle Scholar
  20. 20.
    Zhang L, Tang F, Terracciano L, Hynx D, Kohler R, Bichet S, Hess D, Cron P, Hemmings BA, Hergovich A (2015) NDR functions as a physiological YAP1 kinase in the intestinal epithelium. Curr Biol 25(3):296–305CrossRefGoogle Scholar
  21. 21.
    Cook D, Hoa LY, Gomez V, Gomez M, Hergovich A (2014) Constitutively active NDR1-PIF kinase functions independent of MST1 and hMOB1 signalling. Cell Signal 26(8):1657–1667CrossRefGoogle Scholar
  22. 22.
    Hergovich A, Bichsel SJ, Hemmings BA (2005) Human NDR kinases are rapidly activated by MOB proteins through recruitment to the plasma membrane and phosphorylation. Mol Cell Biol 25(18):8259–8272CrossRefGoogle Scholar
  23. 23.
    Hergovich A, Schmitz D, Hemmings BA (2006) The human tumour suppressor LATS1 is activated by human MOB1 at the membrane. Biochem Biophys Res Commun 345(1):50–58CrossRefGoogle Scholar
  24. 24.
    Hoa L, Kulaberoglu Y, Gundogdu R, Cook D, Mavis M, Gomez M, Gomez V, Hergovich A (2016) The characterisation of LATS2 kinase regulation in Hippo-YAP signalling. Cell Signal 28(5):488–497. CrossRefPubMedGoogle Scholar
  25. 25.
    Kohler RS, Schmitz D, Cornils H, Hemmings BA, Hergovich A (2010) Differential NDR/LATS interactions with the human MOB family reveal a negative role for human MOB2 in the regulation of human NDR kinases. Mol Cell Biol 30(18):4507–4520CrossRefGoogle Scholar
  26. 26.
    Vichalkovski A, Gresko E, Cornils H, Hergovich A, Schmitz D, Hemmings BA (2008) NDR kinase is activated by RASSF1A/MST1 in response to Fas receptor stimulation and promotes apoptosis. Curr Biol 18(23):1889–1895CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Cancer InstituteUniversity College LondonLondonUK

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