Editing of the GluR-B ion channel RNA in vitro by recombinant double-stranded RNA adenosine deaminase
Editing of the GluR-B ion channel RNA in vitro by recombinant double-stranded RNA adenosine deaminase
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DOI：
10.1002/j.1460-2075.1996.tb00331.x
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发表时间：
1996-01-02
期刊：
影响因子：
11.4
通讯作者：
Nishikura, K
中科院分区：
文献类型：
--
作者：
Dabiri, GA;Lai, F;Nishikura, K
双链RNA（dsRNA）特异性腺苷脱氨酶（DRDA）被认为是负责编辑脑中编码谷氨酸门控离子通道亚基（GluR）的RNA转录物的酶。在一种情况下，编辑将基因编码的谷氨酰胺（Q）改变为位于α-氨基-3-羟基-5-甲基异恶唑-4-丙酸（AMPA）通道形成结构域内的精氨酸（R）在该位点（称为“Q/R”位点）编辑的结果是GluR通道的Ca 2+渗透性的显著改变。使用重组表达的DRDA蛋白，我们现在在体外证明DRDA确实参与GluR-B RNA的编辑。除了涉及外显子和内含子序列的RNA双链体结构的形成之外，通过DRDA进行的Q/R位点选择性编辑还需要即使在非神经元细胞中也通常存在的辅因子蛋白。该RNA编辑系统的准确性和效率似乎由DRDA、辅因子和底物GluR-B RNA之间的定量平衡决定。
Double-stranded RNA (dsRNA)-specific adenosine deaminase (DRADA) has been implicated as an enzyme responsible for the editing of RNA transcripts encoding glutamate-gated ion channel subunits (GluR) in brain, In one case, the editing alters the gene-encoded glutamine (Q) to an arginine (R) located within the channel-forming domain of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor subunit GluR-B, The result of editing at this site, called the 'Q/R' site, is a profound alteration of the Ca2+ permeability of the GluR channel, Using recombinantly expressed DRADA proteins, we now demonstrate in vitro that DRADA is indeed involved in editing of the GluR-B RNA. In addition to the formation of an RNA duplex structure involving exon and intron sequences, Q/R site-selective editing by DRADA also requires a cofactor protein(s) commonly present even in non-neuronal cells, The accuracy and efficiency of this RNA editing system appear to be determined by the quantitative balance between DRADA, cofactor and substrate GluR-B RNA.