Journal Article

Functional analysis of rare variants found in schizophrenia implicates a critical role for GIT1–PAK3 signaling in neuroplasticity

Zhang, Q, Rudenko, G, Biag, J, Gangwar, S P, Fleishman, M, Haggarty, S J, Purcell, S M, Fass, D M, Premont, R T, Lewis, M C, Kim, M J, Fromer, M, McCarroll, S A, Machius, M, Scolnick, E M

2017

Overview

Although the pathogenesis of schizophrenia (SCZ) is proposed to involve alterations of neural circuits via synaptic dysfunction, the underlying molecular mechanisms remain poorly understood. Recent exome sequencing studies of SCZ have uncovered numerous single-nucleotide variants (SNVs); however, the majority of these SNVs have unknown functional consequences, leaving their disease relevance uncertain. Filling this knowledge gap requires systematic application of quantitative and scalable assays to assess known and novel biological functions of genes. Here we demonstrate loss-of-function effects of multiple rare coding SNVs found in SCZ subjects in the GIT1 ( G protein-coupled receptor kinase interacting ArfGAP 1 ) gene using functional cell-based assays involving coexpression of GIT1 and PAK3 (p21 protein (Cdc42/Rac)-activated kinase 3). Most notably, a GIT1-R283W variant reported in four independent SCZ cases was defective in activating PAK3 as well as MAPK (mitogen-activated protein kinase). Similar functional deficits were found for a de novo SCZ variant GIT1-S601N. Additional assays revealed deficits in the capacity of GIT1-R283W to stimulate PAK phosphorylation in cultured hippocampal neurons. In addition, GIT1-R283W showed deficits in the induction of GAD1 (glutamate decarboxylase 1) protein expression. Extending these functional assays to 10 additional rare GIT1 variants revealed the existence of an allelic series with the majority of the SCZ case variants exhibiting loss of function toward MAPK activation in a manner correlated with loss of PAK3 activation. Taken together, we propose that rare variants in GIT1, along with other genetic and environmental factors, cause dysregulation of PAK3 leading to synaptic deficits in SCZ.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 13/89 / 631/378 / 692/699/476/1799 / 96/1 / 96/106 / 96/109 / 96/95 / Adaptor Proteins, Signal Transducing - genetics / Adaptor Proteins, Signal Transducing - metabolism / Behavioral Sciences / Biological Psychology / Cdc42 protein / Cell Culture Techniques - methods / Cell Cycle Proteins - genetics / Cell Cycle Proteins - metabolism / Development and progression / Environmental factors / G protein-coupled receptor kinase / Genes / Genetic Variation - genetics / Glutamate decarboxylase / GTP-binding protein / GTPase-Activating Proteins - genetics / Guanine Nucleotide Exchange Factors - metabolism / Health aspects / HEK293 Cells - metabolism / Hippocampus / Hippocampus - metabolism / Hospitals / Humans / Kinases / MAP kinase / Medicine / Medicine & Public Health / Mental disorders / Mitogen-Activated Protein Kinases - metabolism / Molecular modelling / Neural coding / Neural networks / Neurobiology / Neurology / Neuronal Plasticity - genetics / Neuronal Plasticity - physiology / Neurons - metabolism / Neuroplasticity / Neurosciences / original-article / p21-Activated Kinases - genetics / p21-Activated Kinases - metabolism / Pharmacotherapy / Phosphoproteins / Phosphorylation / Polymorphism, Single Nucleotide - genetics / Protein kinase / Protein-Serine-Threonine Kinases - metabolism / Proteins / Psychiatry / Schizophrenia / Schizophrenia - genetics / Signal Transduction - genetics / Signaling peptides and proteins