Novel recognition motifs and biological functions of the RNA-binding protein HuD revealed by genome-wide identification of its targets.
Novel recognition motifs and biological functions of the RNA-binding protein HuD revealed by genome-wide identification of its targets.
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DOI：
10.1093/nar/gkp863
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发表时间：
2010-01
影响因子：
14.9
通讯作者：
Perrone-Bizzozero NI
中科院分区：
文献类型：
--
作者：
Bolognani F;Contente-Cuomo T;Perrone-Bizzozero NI
HUD是一种神经元ELAV样RNA结合蛋白(RBP)，参与神经系统的发育、再生以及学习和记忆。该蛋白通过与3‘端非翻译区富含AU的不稳定元件(ARE)结合来稳定mRNAs。为了分离其体内靶点，首先从脑提取液中免疫沉淀含有HUD的信使核糖核蛋白(MRNP)复合体，并通过随后的GST-HUD下拉和芯片分析鉴定其直接结合的mRNAs。利用最富集靶标的3‘非编码区序列和HUD Are结合位点的已知序列限制，我们发现了三个新的识别基序。基序2和3是富U的，而基序1是富C的。体外结合分析表明，HUD与基序3的亲和力最高，其次是基序2和1，亲和力较小。这些基序被发现在HUD过度表达的小鼠的大脑mRNA中过度表达，支持这些序列的生物学功能。基因本体论分析表明，HUD靶标富含参与神经元分化的信号通路，其中许多mRNAs编码其他限制性商业惯例、翻译因子和肌动蛋白结合蛋白。这些发现为进一步了解HUD促进神经发育和突触可塑性的转录后机制提供了进一步的见解。
HuD is a neuronal ELAV-like RNA-binding protein (RBP) involved in nervous system development, regeneration, and learning and memory. This protein stabilizes mRNAs by binding to AU-rich instability elements (AREs) in their 3′ unstranslated regions (3′ UTR). To isolate its in vivo targets, messenger ribonucleoprotein (mRNP) complexes containing HuD were first immunoprecipitated from brain extracts and directly bound mRNAs identified by subsequent GST-HuD pull downs and microarray assays. Using the 3′ UTR sequences of the most enriched targets and the known sequence restrictions of the HuD ARE-binding site, we discovered three novel recognition motifs. Motifs 2 and 3 are U-rich whereas motif 1 is C-rich. In vitro binding assays indicated that HuD binds motif 3 with the highest affinity, followed by motifs 2 and 1, with less affinity. These motifs were found to be over-represented in brain mRNAs that are upregulated in HuD overexpressor mice, supporting the biological function of these sequences. Gene ontology analyses revealed that HuD targets are enriched in signaling pathways involved in neuronal differentiation and that many of these mRNAs encode other RBPs, translation factors and actin-binding proteins. These findings provide further insights into the post-transcriptional mechanisms by which HuD promotes neural development and synaptic plasticity.
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影响因子：
14.9
作者：
Bakheet T;Williams BR;Khabar KS
通讯作者：
Khabar KS
DOI：
10.1006/bbrc.1998.8247
发表时间：
1998-03-06
影响因子：
3.1
作者：
Dobashi, Y;Shoji, M;Kameya, T
通讯作者：
Kameya, T
影响因子：
4.8
作者：
Beckel-Mitchener, AC;Miera, A;Perrone-Bizzozero, NI
通讯作者：
Perrone-Bizzozero, NI
影响因子：
8
作者：
Halbeisen, R. E.;Galgano, A.;Scherrer, T.;Gerber, A. P.
通讯作者：
Gerber, A. P.
影响因子：
5.3
作者：
Anderson, KD;Sengupta, J;Perrone-Bizzozero, NI
通讯作者：
Perrone-Bizzozero, NI