Isolation and characterization of Dcp1p, the yeast mRNA decapping enzyme
Isolation and characterization of Dcp1p, the yeast mRNA decapping enzyme
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DOI：
10.1093/emboj/17.5.1487
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发表时间：
1998-03-02
期刊：
影响因子：
11.4
通讯作者：
Parker, R
中科院分区：
文献类型：
--
作者：
LaGrandeur, TE;Parker, R
mRNA降解的主要机制是通过去腺苷化、去帽和5' -> 3'外切核酸降解过程发生。最近，DCP 1基因的产物已被证明是体内去帽mRNAs所需的，并与体外去帽活性共纯化。我们已将DCP 1 p纯化至均一，并表明它足以用于去帽，从而表明DCP 1 p是去帽酶，Dcp 1 p的体外活性表征表明，帽结构的7-甲基基团有助于酶的底物特异性，此外，Dcp 1 p被未加帽的mRNA有效地抑制，并且该酶有效地切割长度大于或等于25个核苷酸的底物，优选更长的mRNA底物。这些性质表明Dcp 1 p通过与帽和RNA部分的相互作用识别mRNA底物。Dcp 1 p也是一种磷蛋白，表明其活性可能受转录后修饰的调节。
A major mechanism of mRNA decay occurs by the process of deadenylation, decapping and 5' --> 3' exonucleolytic degradation, Recently, the product of the DCP1 gene has been shown to be required for decapping mRNAs in vivo and co-purifies with decapping activity in vitro, We have purified Dcp1p to homogeneity and shown that it is sufficient for decapping, thereby indicating that Dcp1p is the decapping enzyme, Characterization of Dcp1p activity in vitro indicated that the 7-methyl group off the cap structure contributes to the enzyme's substrate specificity, In addition, Dcp1p was effectively inhibited by uncapped mRNAs, and the enzyme efficiently cleaved substrates that were greater than or equal to 25 nucleotides in length, with a preference for longer mRNA substrates. These properties suggest that Dcp1p recognizes the mRNA substrate by interactions with both the cap and the RNA moiety, The Dcp1p is also a phosphoprotein, suggesting its activity may be regulated by post-transcriptional modification.