Complex formation of the neuron-specific ELAV-like hu RNA-binding proteins
Complex formation of the neuron-specific ELAV-like hu RNA-binding proteins
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DOI：
10.1093/nar/gkf567
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发表时间：
2002-10-15
影响因子：
14.9
通讯作者：
Sakamoto, H
中科院分区：
文献类型：
--
作者：
Kasashima, K;Sakashita, E;Sakamoto, H
Hu蛋白是果蝇ELAV的脊椎动物同源物的RNA结合蛋白，并且涉及在3 '-非翻译区中具有富含AU元件（战神）的特定mRNA的稳定或增强的翻译。在这里，使用酵母双杂交系统，我们表明，神经元特异性胡蛋白可以相互作用。免疫沉淀分析表明，胡蛋白之间的相互作用发生在哺乳动物细胞中，并强烈增强细胞RNA的存在下。此外，使用原位化学交联分析，我们发现，HuD，神经元特异性胡蛋白之一，在细胞中多聚化。交联的HuD多聚体保留特异性RNA结合能力，并且可以与另外的Hu蛋白相互作用。与这种生化特性一致，HuD在两种神经源性细胞系中表现出颗粒状分布。这些结果表明，RNA结合形式的HuD多聚化合作，形成一个特定的颗粒状结构，可以作为一个网站的转录后调控ARE的mRNA。
Hu proteins are RNA-binding proteins that are the vertebrate homologs of Drosophila ELAV, and are implicated in stabilization or enhanced translation of specific mRNAs with AU-rich elements (AREs) in the 3'-untranslated region. Here, using the yeast two-hybrid system, we show that neuron-specific Hu proteins can interact with themselves. Immuno precipitation assays demonstrated that the interaction between Hu proteins occurs in mammalian cells and is strongly enhanced in the presence of cellular RNA. Furthermore, using in situ chemical crosslinking assays, we found that HuD, one of the neuron-specific Hu proteins, multimerizes in cells. The crosslinked HuD multimers retain specific RNA-binding ability and can interact with additional Hu proteins. Consistent with this biochemical property, HuD showed granular distribution in two neurogenic cell lines. These results suggest that the RNA-bound form of HuD multimerizes cooperatively to form a specific granular structure that may serve as a site of post-transcriptional regulation of ARE-containing mRNAs.