Nova regulates brain-specific splicing to shape the synapse
Nova regulates brain-specific splicing to shape the synapse
复制标题
DOI：
10.1038/ng1610
复制
发表时间：
2005-08-01
期刊：
影响因子：
30.8
通讯作者：
Darnell, RB
中科院分区：
文献类型：
--
作者：
Ule, J;Ule, A;Darnell, RB
替代RNA剪接大大增加了蛋白质组的多样性，因此可能有助于组织特异性功能。我们使用自定义Affymetrix微阵列对替代剪接进行了全基因组定量分析，以评估神经元素剪接因子NOVA在大脑中的作用。我们使用严格的算法来鉴定591个外显子，这些外显子相对于免疫组织在大脑中差异剪接，其中6.6％显示了Nova2（ - / - ）小鼠新皮层的主要剪接缺陷。我们测试了49个外显子，其中最大的预测NOVA依赖性剪接变化，并通过RT-PCR验证了所有49个外显子。我们分析了编码的蛋白质，发现所有具有确定脑功能的人都作用在突触中（40个中的34个，包括神经递质受体，阳离子通道，粘附和支架蛋白）或轴突指导（40中的8个）。此外，在具有已知相互作用伙伴的35种蛋白质中，有74％（26）相互作用。验证大量的NOVA RNA目标使我们确定了一个多层网络，其中NOVA调节编码在突触中相互作用的RNA的外显子含量。
Alternative RNA splicing greatly increases proteome diversity and may thereby contribute to tissue-specific functions. We carried out genome-wide quantitative analysis of alternative splicing using a custom Affymetrix microarray to assess the role of the neuronal splicing factor Nova in the brain. We used a stringent algorithm to identify 591 exons that were differentially spliced in the brain relative to immune tissues, and 6.6% of these showed major splicing defects in the neocortex of Nova2(-/-) mice. We tested 49 exons with the largest predicted Nova-dependent splicing changes and validated all 49 by RT-PCR. We analyzed the encoded proteins and found that all those with defined brain functions acted in the synapse (34 of 40, including neurotransmitter receptors, cation channels, adhesion and scaffold proteins) or in axon guidance ( 8 of 40). Moreover, of the 35 proteins with known interaction partners, 74% (26) interact with each other. Validating a large set of Nova RNA targets has led us to identify a multi-tiered network in which Nova regulates the exon content of RNAs encoding proteins that interact in the synapse.