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A kinome-wide RNAi screen identifies ERK2 as a druggable regulator of Shank3 stability

Molecular Psychiatry volume 25, pages 2504–2516 (2020)Cite this article

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Abstract

Neurons are sensitive to changes in the dosage of many genes, especially those regulating synaptic functions. Haploinsufficiency of SHANK3 causes Phelan-McDermid syndrome and autism, whereas duplication of the same gene leads to SHANK3 duplication syndrome, a disorder characterized by neuropsychiatric phenotypes including hyperactivity and bipolar disorder as well as epilepsy. We recently demonstrated the functional modularity of Shank3, which suggests that normalizing levels of Shank3 itself might be more fruitful than correcting pathways that function downstream of it for treatment of disorders caused by alterations in SHANK3 dosage. To identify upstream regulators of Shank3 abundance, we performed a kinome-wide siRNA screen and identified multiple kinases that potentially regulate Shank3 protein stability. Interestingly, we discovered that several kinases in the MEK/ERK2 pathway destabilize Shank3 and that genetic deletion and pharmacological inhibition of ERK2 increases Shank3 abundance in vivo. Mechanistically, we show that ERK2 binds Shank3 and phosphorylates it at three residues to promote its poly-ubiquitination-dependent degradation. Altogether, our findings uncover a druggable pathway as a potential therapeutic target for disorders with reduced SHANK3 dosage, provide a rich resource for studying Shank3 regulation, and demonstrate the feasibility of this approach for identifying regulators of dosage-sensitive genes.

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Acknowledgements

We are grateful to Zhou Songyang, Dan Liu, and Jun Xu for providing plasmids and help for the BiFC assay; Maxime WC Rousseaux, Nan Lu, Laura Lombardi, Paymaan Jafar-Nejad, Jeehye Park, and all Zoghbi lab members for technical assistance and insightful discussions on this work. This work was supported by the Howard Hughes Medical Institute (HYZ), a gift from Mr. Charif Souki (to the Jan and Dan Duncan Neurological Research Institute), Autism Speaks (grant #9120 to LW), NINDS K08 Award NS091381 (JLH), the Robbins Foundation (JLH), Baylor Pediatrics Pilot Award (JLH), Chao Physician-Scientist Award (JLH), the Cytometry and Cell Sorting Core at Baylor College of Medicine with funding from the NIH (P30 AI036211, P30 CA125123, and S10 RR024574) and the expert assistance of Joel M. Sederstrom.

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Authors and Affiliations

  1. Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, 77030, USA

    Li Wang, Carolyn J. Adamski, Vitaliy V. Bondar, Evelyn Craigen, Kihoon Han & Huda Y. Zoghbi

  2. Jan and Dan Duncan Neurological Research Institute at Texas Children’s Hospital, Houston, TX, 77030, USA

    Li Wang, Carolyn J. Adamski, Vitaliy V. Bondar, Evelyn Craigen, Kaifang Pang, Kihoon Han, Zhandong Liu, J. Lloyd Holder Jr. & Huda Y. Zoghbi

  3. Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX, 77030, USA

    Carolyn J. Adamski & Huda Y. Zoghbi

  4. Department of Pathology & Immunology, Baylor College of Medicine, Houston, TX, 77030, USA

    John R. Collette & Richard N. Sifers

  5. Department of Pediatrics, Baylor College of Medicine, Houston, TX, 77030, USA

    Kaifang Pang, Zhandong Liu, J. Lloyd Holder Jr. & Huda Y. Zoghbi

  6. Department of Neuroscience and Division of Brain Korea 21 Biomedical Science, Korea University College of Medicine, Seoul, 02841, South Korea

    Kihoon Han

  7. Alkek Center for Molecular Discovery, Verna and Marrs McLean, Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA

    Antrix Jain & Sung Y. Jung

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  1. Li Wang
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Correspondence to J. Lloyd Holder Jr. or Huda Y. Zoghbi.

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Wang, L., Adamski, C.J., Bondar, V.V. et al. A kinome-wide RNAi screen identifies ERK2 as a druggable regulator of Shank3 stability. Mol Psychiatry 25, 2504–2516 (2020). https://doi.org/10.1038/s41380-018-0325-9

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