Laboratory of Molecular Neuro-Oncology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Neuron. 2012 Sep 20;75(6):1067-80. doi: 10.1016/j.neuron.2012.07.009.
The paraneoplastic neurologic disorders target several families of neuron-specific RNA binding proteins (RNABPs), revealing that there are unique aspects of gene expression regulation in the mammalian brain. Here, we used HITS-CLIP to determine robust binding sites targeted by the neuronal Elav-like (nElavl) RNABPs. Surprisingly, nElav protein binds preferentially to GU-rich sequences in vivo and in vitro, with secondary binding to AU-rich sequences. nElavl null mice were used to validate the consequence of these binding events in the brain, demonstrating that they bind intronic sequences in a position dependent manner to regulate alternative splicing and to 3'UTR sequences to regulate mRNA levels. These controls converge on the glutamate synthesis pathway in neurons; nElavl proteins are required to maintain neurotransmitter glutamate levels, and the lack of nElavl leads to spontaneous epileptic seizure activity. The genome-wide analysis of nElavl targets reveals that one function of neuron-specific RNABPs is to control excitation-inhibition balance in the brain.
副肿瘤性神经紊乱靶向几种神经元特异性 RNA 结合蛋白 (RNABPs) 家族，揭示了哺乳动物大脑中基因表达调控的独特方面。在这里，我们使用 HITS-CLIP 来确定神经元 Elav 样 (nElavl) RNABPs 的稳健结合位点。令人惊讶的是，nElav 蛋白在体内和体外优先结合富含 GU 的序列，其次是富含 AU 的序列。nElavl 缺失小鼠被用于验证这些结合事件在大脑中的后果，证明它们以依赖位置的方式结合内含子序列以调节选择性剪接，并结合 3'UTR 序列以调节 mRNA 水平。这些控制在神经元中的谷氨酸合成途径中汇聚；nElavl 蛋白对于维持神经递质谷氨酸水平是必需的，缺乏 nElavl 会导致自发性癫痫发作活动。nElavl 靶点的全基因组分析表明，神经元特异性 RNABPs 的一个功能是控制大脑中的兴奋-抑制平衡。
