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Autopalmitoylation of TEAD proteins regulates transcriptional output of the Hippo pathway

Nature Chemical Biology volume 12, pages 282–289 (2016)Cite this article

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Abstract

TEA domain (TEAD) transcription factors bind to the coactivators YAP and TAZ and regulate the transcriptional output of the Hippo pathway, playing critical roles in organ size control and tumorigenesis. Protein S-palmitoylation attaches a fatty acid, palmitate, to cysteine residues and regulates protein trafficking, membrane localization and signaling activities. Using activity-based chemical probes, we discovered that human TEADs possess intrinsic palmitoylating enzyme–like activities and undergo autopalmitoylation at evolutionarily conserved cysteine residues under physiological conditions. We determined the crystal structures of lipid-bound TEADs and found that the lipid chain of palmitate inserts into a conserved deep hydrophobic pocket. Strikingly, palmitoylation did not alter TEAD's localization, but it was required for TEAD's binding to YAP and TAZ and was dispensable for its binding to the Vgll4 tumor suppressor. Moreover, palmitoylation-deficient TEAD mutants impaired TAZ-mediated muscle differentiation in vitro and tissue overgrowth mediated by the Drosophila YAP homolog Yorkie in vivo. Our study directly links autopalmitoylation to the transcriptional regulation of the Hippo pathway.

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Figure 1: Chemical approaches reveal that TEA domain (TEAD) transcription factors are palmitoylated.
Figure 2: TEAD is autopalmitoylated at evolutionarily conserved cysteine residues under physiological palmitoyl-CoA concentrations.
Figure 3: Structures of palmitate-bound human TEAD2 YBD and TEAD1–YAP complex.
Figure 4: Palmitoylation of TEAD is required for its association with YAP/TAZ.
Figure 5: Palmitoylation regulates TEAD functions in muscle cell differentiation in vitro.
Figure 6: Palmitoylation is required for the functions of Drosophila TEAD protein Scalloped (Sd) in vivo.

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Acknowledgements

This work was supported by a Stewart Rahr–MRA (Melanoma Research Alliance) Young Investigator Award, a Department of Defense (DoD) Career Development Award (W81XWH-13-1-0203), and grants from the American Cancer Society (124929-RSG-13-291-01-TBE), US National Institutes of Health/National Cancer Institute (R01CA181537) (to X.W.), National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Diseases (R01DK107651-01) (to X.W. and X.L.), National Institutes of Health/National Institute of General Medical Sciences (R01GM107415) (to X.L.) and National Institute of Health/National Eye Institute (R01EY015708) (to D.P.); D.P. is supported by the Howard Hughes Medical Institute. We thank N. Bardeesy, M. Fukata, K.-L. Guan and K. White for constructs and cell lines; J.-R.J. Yeh, H. Yu and N. Gray for discussion and critical comments on the manuscript; and the Taplin Mass Spec Core facility at Harvard Medical School and the Proteomics Core at UTSW for proteomic studies. Use of the Argonne National Laboratory Structural Biology Center beamlines at the Advanced Photon Source was supported by the US Department of Energy (DOE) under contract DE-AC02-06CH11357.

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  1. PuiYee Chan and Xiao Han: These authors contributed equally to this work.

Authors and Affiliations

  1. Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts, USA

    PuiYee Chan, Baohui Zheng, Michael DeRan, Gopala K Jarugumilli & Xu Wu

  2. Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, School of Life Sciences, Jilin University, Changchun, China

    Xiao Han

  3. Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Xiao Han & Xuelian Luo

  4. Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Xiao Han & Xuelian Luo

  5. Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Jianzhong Yu, Hua Deng & Duojia Pan

  6. Department of Molecular Biology & Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Jianzhong Yu, Hua Deng & Duojia Pan

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Contributions

X.W. conceived the concepts, designed the experiments and supervised the studies. P.C. designed and performed the cell biology and biochemistry experiments with the help of M.D. X.H. performed protein purification, crystallization and structure determination and carried out MS analysis of TEAD2 autopalmitoylation. B.Z. and G.K.J. synthesized the probes. B.Z. identified TEAD from mass spec studies and tested the DHHC family of PATs. J.Y., H.D. and D.P. carried out Drosophila genetics experiments. X.L. contributed to experimental design and structure refinements of palmitate-bound TEAD2 and TEAD1–YAP complex. P.C., D.P., X.L. and X.W. analyzed the data; P.C., D.P. X.L. and X.W. wrote the manuscript with input from all coauthors.

Corresponding authors

Correspondence to Xuelian Luo or Xu Wu.

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Chan, P., Han, X., Zheng, B. et al. Autopalmitoylation of TEAD proteins regulates transcriptional output of the Hippo pathway. Nat Chem Biol 12, 282–289 (2016). https://doi.org/10.1038/nchembio.2036

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