NeuN/Rbfox3 nuclear and cytoplasmic isoforms differentially regulate alternative splicing and nonsense-mediated decay of Rbfox2.
NeuN/Rbfox3 nuclear and cytoplasmic isoforms differentially regulate alternative splicing and nonsense-mediated decay of Rbfox2.
复制标题
DOI：
10.1371/journal.pone.0021585
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Jensen KB
中科院分区：
文献类型：
--
作者：
Dredge BK;Jensen KB
抗NeuN（神经元核）是一种广泛用于特异性检测有丝分裂后神经元的单克隆抗体。抗NeuN反应性主要是核反应性;通过Western检测，其检测到分子量范围为45 kDa至>75 kDa的多个条带。表达筛选质粒鉴定R3 hdm 2为NeuN;然而，在45-50 kDa处的两种主要NeuN种类的免疫沉淀和质谱鉴定两者均为RNA结合蛋白Rbfox 3（Fox家族的选择性剪接因子的成员），确认并扩展了Kim等人将45 kDa条带鉴定为Rbfox 3。R3 hdm 2和Rbfox 3中的NeuN反应性表位揭示了位于Rbfox 3蛋白的N-末端的共同的富含脯氨酸和谷氨酰胺的结构域。我们的数据表明，选择性剪接的Rbfox 3前体mRNA本身导致产生四种蛋白质亚型，迁移在45-50 kDa的范围内，这些剪接的选择之一，调节Rbfox 3/NeuN亚细胞稳态分布，通过添加或删除一个短的C-末端延伸含有二分疏水性脯氨酸-酪氨酸核定位信号的第二部分。Rbfox 3调节Rbfox 2前mRNA的选择性剪接，产生编码Rbfox 2蛋白显性负性形式的信息。在这里，我们表明，核Rbfox 3亚型也可以增强包含在Rbfox 2 mRNA的隐蔽外显子，导致无义介导的衰减的消息，从而有助于通过一种新的机制Rbfox 3的负调控Rbfox 2。
Anti-NeuN (Neuronal Nuclei) is a monoclonal antibody used extensively to specifically detect post-mitotic neurons. Anti-NeuN reactivity is predominantly nuclear; by western it detects multiple bands ranging in molecular weight from 45 kDa to >75 kDa. Expression screening putatively identified R3hdm2 as NeuN; however immunoprecipitation and mass spectrometry of the two major NeuN species at 45–50 kDa identified both as the RNA binding protein Rbfox3 (a member of the Fox family of alternative splicing factors), confirming and extending the identification of the 45 kDa band as Rbfox3 by Kim et al. Mapping of the anti-NeuN reactive epitopes in both R3hdm2 and Rbfox3 reveals a common proline- and glutamine-rich domain that lies at the N-terminus of the Rbfox3 protein. Our data suggests that alternative splicing of the Rbfox3 pre-mRNA itself leads to the production of four protein isoforms that migrate in the 45–50 kDa range, and that one of these splicing choices regulates Rbfox3/NeuN sub-cellular steady-state distribution, through the addition or removal of a short C-terminal extension containing the second half of a bipartite hydrophobic proline-tyrosine nuclear localization signal. Rbfox3 regulates alternative splicing of the Rbfox2 pre-mRNA, producing a message encoding a dominant negative form of the Rbfox2 protein. We show here that nuclear Rbfox3 isoforms can also enhance the inclusion of cryptic exons in the Rbfox2 mRNA, resulting in nonsense-mediated decay of the message, thereby contributing to the negative regulation of Rbfox2 by Rbfox3 through a novel mechanism.
登录
查看更多内容
影响因子：
10.5
作者：
Ni, Julie Z.;Grate, Leslie;Ares, Manuel, Jr.
通讯作者：
Ares, Manuel, Jr.
影响因子：
64.5
作者：
Fornerod, M;Ohno, M;Mattaj, IW
通讯作者：
Mattaj, IW
影响因子：
14.9
作者：
通讯作者：
--
影响因子：
4.8
作者：
Ladd, AN;Nguyen, NH;Cooper, TA
通讯作者：
Cooper, TA
影响因子：
4.5
作者：
Damianov, Andrey;Black, Douglas L.
通讯作者：
Black, Douglas L.