Lai F, Chen C X, Carter K C, Nishikura K
The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Mol Cell Biol. 1997 May;17(5):2413-24. doi: 10.1128/MCB.17.5.2413.
Double-stranded (ds) RNA-specific adenosine deaminase converts adenosine residues into inosines in dsRNA and edits transcripts of certain cellular and viral genes such as glutamate receptor (GluR) subunits and hepatitis delta antigen. The first member of this type of deaminase, DRADA1, has been recently cloned based on the amino acid sequence information derived from biochemically purified proteins. Our search for DRADA1-like genes through expressed sequence tag databases led to the cloning of the second member of this class of enzyme, DRADA2, which has a high degree of sequence homology to DRADA1 yet exhibits a distinctive RNA editing site selectivity. There are four differentially spliced isoforms of human DRADA2. These different isoforms of recombinant DRADA2 proteins, including one which is a human homolog of the recently reported rat RED1, were analyzed in vitro for their GluR B subunit (GluR-B) RNA editing site selectivity. As originally reported for rat RED1, the DRADA2a and -2b isoforms edit GluR-B RNA efficiently at the so-called Q/R site, whereas DRADA1 barely edits this site. In contrast, the R/G site of GluR-B RNA was edited efficiently by the DRADA2a and -2b isoforms as well as DRADA1. Isoforms DRADA2c and -2d, which have a distinctive truncated shorter C-terminal structure, displayed weak adenosine-to-inosine conversion activity but no editing activity tested at three known sites of GluR-B RNA. The possible role of these DRADA2c and -2d isoforms in the regulatory mechanism of RNA editing is discussed.
双链(ds)RNA特异性腺苷脱氨酶可将dsRNA中的腺苷残基转化为肌苷,并对某些细胞和病毒基因的转录本进行编辑,如谷氨酸受体(GluR)亚基和丁型肝炎抗原。这类脱氨酶的首个成员DRADA1,最近已根据从生化纯化蛋白中获得的氨基酸序列信息进行了克隆。我们通过表达序列标签数据库搜索DRADA1样基因,从而克隆出了这类酶的第二个成员DRADA2,它与DRADA1具有高度的序列同源性,但表现出独特的RNA编辑位点选择性。人DRADA2有四种不同的剪接异构体。对这些重组DRADA2蛋白的不同异构体,包括一种是最近报道的大鼠RED1的人类同源物,在体外分析了它们对GluR B亚基(GluR-B)RNA编辑位点的选择性。正如最初对大鼠RED1的报道,DRADA2a和-2b异构体在所谓的Q/R位点高效编辑GluR-B RNA,而DRADA1几乎不编辑该位点。相反,GluR-B RNA的R/G位点被DRADA2a和-2b异构体以及DRADA1高效编辑。具有独特截短的较短C末端结构的DRADA2c和-2d异构体,显示出较弱的腺苷到肌苷的转化活性,但在GluR-B RNA的三个已知位点测试时没有编辑活性。讨论了这些DRADA2c和-2d异构体在RNA编辑调控机制中的可能作用。