Uridine Insertion/Deletion RNA Editing in Trypanosomatids: Specific Stimulation in vitro of Leishmania tarentolae REL1 RNA Ligase Activity by the MP63 Zinc Finger Protein
Uridine Insertion/Deletion RNA Editing in Trypanosomatids: Specific Stimulation in vitro of Leishmania tarentolae REL1 RNA Ligase Activity by the MP63 Zinc Finger Protein
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DOI：
10.1016/j.protis.2010.01.001
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发表时间：
2010-07-01
期刊：
影响因子：
2.5
通讯作者：
Simpson, Larry
中科院分区：
文献类型：
--
作者：
Gao, Guanghan;Rogers, Kestrel;Simpson, Larry
锥虫线粒体中线粒体 mRNA 的 U 插入/删除 RNA 编辑是由核心复合物 (RECC) 介导的，该核心复合物包含约 16-20 个蛋白质，该蛋白质通过 RNA 与其他几个多蛋白质复合物连接。 RECC 中有两个已知的子复合体：REL1 子复合体，其中包含 REL1 RNA 连接酶、MP63 锌指蛋白和 REX2U 特异性 3'-5' 核酸外切酶； REL2 子复合体包含 REL2 RNA 连接酶、RET2 3' TUTase 和 MP81 锌指蛋白。在这项研究中，我们在杆状病毒感染的昆虫细胞中表达后，亲和分离了带有 TAP 标签的重组利什曼原虫 RET2 和塔伦托利什曼原虫 MP63、REL1 和 REL2 蛋白。研究发现重组 MP63 蛋白可刺激重组 REL1 的多种体外活性；这些活动包括自腺苷酸化、桥接，甚至使用 REL2 子复合体中的 RET2 进行预切割 gRNA 介导的 U 插入编辑。重组 MP63 对类似的 REL2 连接活性没有影响。 REL1 的特异性与 MP63 作为 REL1 子复合物的组成部分一致。这些结果表明，体内 MP63 与 REL1 的相互作用可能在调节 RNA 编辑的整体活性中发挥作用。 (C) 2010 爱思唯尔有限公司。版权所有。
U-insertion/deletion RNA editing of mitochondrial mRNAs in trypanosome mitochondria is mediated by a core complex (RECC) containing around 16-20 proteins which is linked to several other multiprotein complexes by RNA. There are two known subcomplexes in the RECC: the REL1 subcomplex which contains the REL1 RNA ligase, the MP63 zinc finger-containing protein and the REX2U-specific 3'-5' exonuclease; and the REL2 subcomplex which contains the REL2 RNA ligase, the RET2 3' TUTase and the MP81 zinc finger-containing protein. In this study we have affinity isolated recombinant TAP-tagged Leishmania major RET2 and Leishmania tarentolae MP63, REL1 and REL2 proteins after expression in baculovirus-infected insect cells. Recombinant MP63 protein was found to stimulate several invitro activities of recombinant REL1; these activities include autoadenylation, bridged ligation and even pre-cleaved gRNA-mediated U-insertion editing with RET2 which is in the REL2 subcomplex. There was no effect of recombinant MP63 on similar REL2 ligation activities. The specificity for REL1 is consistent with MP63 being a component of the REL1 subcomplex. These results suggest that in vivo the interaction of MP63 with REL1 may play a role in regulating the overall activity of RNA editing. (C) 2010 Elsevier GmbH. All rights reserved.