Novel calcineurin A (PPP3CA) variant associated with epilepsy, constitutive enzyme activation and downregulation of protein expression

• Published in: European journal of human genetics : EJHG Vol. 27; no. 1; pp. 61 - 69
• Main Authors: Rydzanicz, Małgorzata, Wachowska, Małgorzata, Cook, Erik C., Lisowski, Paweł, Kuźniewska, Bożena, Szymańska, Krystyna, Diecke, Sebastian, Prigione, Alessandro, Szczałuba, Krzysztof, Szybińska, Aleksandra, Koppolu, Agnieszka, Murcia Pienkowski, Victor, Kosińska, Joanna, Wiweger, Małgorzata, Kostrzewa, Grażyna, Brzozowska, Małgorzata, Domańska-Pakieła, Dorota, Jurkiewicz, Elżbieta, Stawiński, Piotr, Gromadka, Agnieszka, Zielenkiewicz, Piotr, Demkow, Urszula, Dziembowska, Magdalena, Kuźnicki, Jacek, Creamer, Trevor P., Płoski, Rafał
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01.01.2019; Springer International Publishing
• Subjects: Calcineurin; Calcineurin - genetics; Calcineurin - metabolism; Calcium-binding protein; Calmodulin; Cells, Cultured; Child; Craniofacial Abnormalities - genetics; Craniofacial Abnormalities - pathology; Down-Regulation; Encephalopathy; Enzymes; Epilepsy; Epilepsy - genetics; Epilepsy - pathology; Gene expression; Humans; Kinases; Lymphocytes; Male; mRNA; Mutation, Missense; Neurodevelopmental disorders; Phenotype; Phenotypes; Protein expression; Protein phosphatase; Proteins; Syndrome
• ISSN: 1018-4813; 1476-5438; 1476-5438
• DOI: 10.1038/s41431-018-0254-8

PPP3CA encodes calmodulin-binding catalytic subunit of calcineurin, a ubiquitously expressed calcium/calmodulin-regulated protein phosphatase. Recently de novo PPP3CA variants were reported as a cause of disease in 12 subjects presenting with epileptic encephalopathy and dysmorphic features. We describe a boy with similar phenotype and severe early onset epileptic encephalopathy in whom a novel de novo c.1324C>T (p.(Gln442Ter)) PPP3CA variant was found by whole exome sequencing. Western blot experiments in patient's cells (EBV transformed lymphocytes and neuronal cells derived through reprogramming) indicate that despite normal mRNA abundance the protein expression level is strongly reduced both for the mutated and wild-type protein. By in vitro studies with recombinant protein expressed in E. coli we show that c.1324C>T (p.(Gln442Ter)) results in constitutive activation of the enzyme. Our results confirm the role of PPP3CA defects in pathogenesis of a distinct neurodevelopmental disorder including severe epilepsy and dysmorphism and provide further functional clues regarding the pathogenic mechanism.