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

• Published in: Molecular psychiatry Vol. 25; no. 10; pp. 2504 - 2516
• Main Authors: Wang, Li, Adamski, Carolyn J., Bondar, Vitaliy V., Craigen, Evelyn, Collette, John R., Pang, Kaifang, Han, Kihoon, Jain, Antrix, Y. Jung, Sung, Liu, Zhandong, Sifers, Richard N., Holder, J. Lloyd, Zoghbi, Huda Y.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.10.2020; Nature Publishing Group
• Subjects: 13/106; 13/31; 13/89; 38/70; 45/47; 631/208; 631/61; 64/110; 692/699/476/1373; 82/58; Animals; Antibiotics; Autism; Behavioral Sciences; Biological Psychology; Bipolar disorder; Cell Line, Tumor; Chromosome Disorders - genetics; Cloning; Drug dosages; Epilepsy; Ethylenediaminetetraacetic acid; Extracellular signal-regulated kinase; Female; Flow cytometry; Gene Deletion; Genes; Genetic screening; Haploinsufficiency; Health aspects; Humans; Hyperactivity; Kinases; Male; Medical schools; Medicine; Medicine & Public Health; Mental disorders; Mice; Mitogen-Activated Protein Kinase 1 - antagonists & inhibitors; Mitogen-Activated Protein Kinase 1 - genetics; Mitogen-Activated Protein Kinase 1 - metabolism; Mutation; Nerve Tissue Proteins - deficiency; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neurons - metabolism; Neurosciences; Pharmacotherapy; Phenotypes; Phosphorylation; Phosphorylation - drug effects; Plasmids; Protein Stability - drug effects; Proteins; Psychiatry; Regulation; RNA Interference; RNA-mediated interference; siRNA; Therapeutic targets; Ubiquitination
• ISSN: 1359-4184; 1476-5578
• DOI: 10.1038/s41380-018-0325-9

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.