Glazov Eugene, Phillips Kenneth, Budziszewski Gregory J, Schöb Hanspeter, Meins Frederick, Levin Joshua Z
Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, Maulbeerstrasse 66, CH-4058 Basel, Switzerland.
Plant J. 2003 Aug;35(3):342-9. doi: 10.1046/j.1365-313x.2003.01810.x.
Post-transcriptional gene silencing (PTGS) and the closely related phenomenon RNA interference (RNAi) result from the initial endonucleolytic cleavage of target mRNAs, which are then presumed to be completely hydrolyzed by exoribonucleases. To date, no plant genes required for PTGS are known to encode exoribonucleases. The Arabidopsis Werner Syndrome-like exonuclease (WEX) gene encodes an RNase D domain most similar to that in human Werner Syndrome protein (WRN), but lacks the RecQ helicase domain. It is also related to Caenorhabditis elegans mut-7, which is essential for RNAi, PTGS, and transposon activity. We isolated a loss-of-function mutant, wex-1, that showed greatly reduced expression of WEX mRNA and early flowering. Although wex-1 did not affect expression of a robust marker for transcriptional gene silencing (TGS), PTGS of a green-fluorescent-protein (GFP) reporter gene was blocked in wex-1 and restored by ectopic expression of WEX, indicating that WEX is required for PTGS but not TGS. Thus, members of the RNase D protein family are required for PTGS in both plants and animals. Interestingly, WEX has been shown to interact with an Arabidopsis RecQ helicase, suggesting that these proteins might comprise a functional equivalent of WRN.
转录后基因沉默(PTGS)以及与之密切相关的现象RNA干扰(RNAi)源于靶mRNA的初始内切核酸酶切割,随后这些mRNA被推测会被外切核糖核酸酶完全水解。迄今为止,尚未发现PTGS所需的植物基因编码外切核糖核酸酶。拟南芥沃纳综合征样外切核酸酶(WEX)基因编码一个与人类沃纳综合征蛋白(WRN)中最相似的RNase D结构域,但缺乏RecQ解旋酶结构域。它还与秀丽隐杆线虫的mut-7相关,mut-7对于RNAi、PTGS和转座子活性至关重要。我们分离出了一个功能缺失突变体wex-1,它显示出WEX mRNA的表达大幅降低且开花提前。尽管wex-1不影响转录基因沉默(TGS)的一个强标记的表达,但绿色荧光蛋白(GFP)报告基因的PTGS在wex-1中被阻断,并通过WEX的异位表达得以恢复,这表明WEX是PTGS所必需的,但不是TGS所必需的。因此,RNase D蛋白家族成员在植物和动物的PTGS中都是必需的。有趣的是,WEX已被证明与拟南芥RecQ解旋酶相互作用,这表明这些蛋白质可能构成了与WRN功能相当的物质。