Polyadenylation site-specific differences in the activity of the neuronal βCstF-64 protein in PC-12 cells.
Polyadenylation site-specific differences in the activity of the neuronal βCstF-64 protein in PC-12 cells.
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DOI：
10.1016/j.gene.2013.08.007
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发表时间：
2013-10-25
期刊：
影响因子：
3.5
通讯作者：
MacDonald CC
中科院分区：
文献类型：
--
作者：
Shankarling GS;MacDonald CC
最近的全基因组分析表明，替代性多聚腺苷酸化-调节mRNA 3′端形成的过程-是影响mRNA代谢多个步骤的关键机制，除了增加基因组的蛋白质编码能力之外。虽然替代多聚腺苷酸化的功能后果是众所周知的，蛋白质的因素，调节这一进程的特点很差。以前，我们描述了一个进化上保守的CstF-64 mRNA神经元剪接变体βCstF-64家族，我们假设它在神经系统中的选择性多聚腺苷酸化中起作用。在本研究中，我们发现，βCstF-64 mRNA和蛋白表达增加响应神经生长因子（NGF），伴随着肾上腺PC-12细胞分化为神经元表型，提示βCstF-64在神经元基因表达中的作用。以PC-12细胞为模型，我们发现βCstF-64是一种真正的多聚腺苷酸化蛋白，这可以通过它与CstF复合物的结合以及它刺激荧光素酶报告基因mRNA多聚腺苷酸化的能力来证明。使用荧光素酶测定，我们表明βCstF-64在β-内收蛋白mRNA的两个弱poly（A）位点同等地刺激多聚腺苷酸化。值得注意的是，我们证明了βCstF-64的活性在强聚腺苷酸化信号下低于CstF-64，表明βCstF-64蛋白的活性存在聚腺苷酸化位点特异性差异。我们的数据首次阐明了βCstF-64的多聚腺苷酸化功能，并为神经系统中选择性poly（A）位点的机制提供了初步见解。
Recent genome-wide analyses have implicated alternative polyadenylation — the process of regulated mRNA 3′ end formation — as a critical mechanism that influences multiple steps of mRNA metabolism in addition to increasing the protein-coding capacity of the genome. Although the functional consequences of alternative polyadenylation are well known, protein factors that regulate this process are poorly characterized. Previously, we described an evolutionarily conserved family of neuronal splice variants of the CstF-64 mRNA, βCstF-64, that we hypothesized to function in alternative polyadenylation in the nervous system. In the present study, we show that βCstF-64 mRNA and protein expression increase in response to nerve growth factor (NGF), concomitant with differentiation of adrenal PC-12 cells into a neuronal phenotype, suggesting a role for βCstF-64 in neuronal gene expression. Using PC-12 cells as model, we show that βCstF-64 is a bona fide polyadenylation protein, as evidenced by its association with the CstF complex, and by its ability to stimulate polyadenylation of luciferase reporter mRNA. Using luciferase assays, we show that βCstF-64 stimulates polyadenylation equivalently at the two weak poly(A) sites of the β-adducin mRNA. Notably, we demonstrate that the activity of βCstF-64 is less than CstF-64 on a strong polyadenylation signal, suggesting polyadenylation site-specific differences in the activity of the βCstF-64 protein. Our data address the polyadenylation functions of βCstF-64 for the first time, and provide initial insights into the mechanism of alternative poly(A) site selection in the nervous system.
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影响因子：
3.7
作者：
Li W;Yeh HJ;Shankarling GS;Ji Z;Tian B;MacDonald CC
通讯作者：
MacDonald CC
影响因子：
16.2
作者：
Flavell, Steven W.;Kim, Tae-Kyung;Gray, Jesse M.;Harmin, David A.;Hemberg, Martin;Hong, Elizabeth J.;Markenscoff-Papadimitriou, Eirene;Bear, Daniel M.;Greenberg, Michael E.
通讯作者：
Greenberg, Michael E.
β-无元素前mRNA的脑特异性启动子和聚腺苷酸位点产生异常长的3'-UTR。
影响因子：
14.9
作者：
Costessi, L;Devescovi, G;Baralle, FE;Muro, AF
通讯作者：
Muro, AF
影响因子：
14.9
作者：
Millevoi S;Vagner S
通讯作者：
Vagner S
影响因子：
3.7
作者：
Metheny, LJ;Skuse, GR
通讯作者：
Skuse, GR