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首页> 外文期刊>Molecular and Cellular Biology >Assembly of mitochondrial ribonucleoprotein complexes involves specific guide RNA (gRNA)-binding proteins and gRNA domains but does not require preedited mRNA.
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Assembly of mitochondrial ribonucleoprotein complexes involves specific guide RNA (gRNA)-binding proteins and gRNA domains but does not require preedited mRNA.

机译：线粒体核糖核蛋白复合物的组装涉及特定的指导RNA（gRNA）结合蛋白和gRNA域，但不需要预先编辑的mRNA。

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RNA editing in kinetoplastids probably employs a macromolecular complex, the editosome, that is likely to include the guide RNAs (gRNAs) which specify the edited sequence. Specific ribonucleoprotein (RNP) complexes which form in vitro with gRNAs (H. U. G?ringer, D. J. Koslowsky, T. H. Morales, and K. D. Stuart, Proc. Natl. Acad. Sci. USA, in press) are potential editosomes or their precursors. We find that several factors are important for in vitro formation of these RNP complexes and identify specific gRNA-binding proteins present in the complexes. Preedited mRNA promotes the in vitro formation of the four major gRNA-containing RNP complexes under some conditions but is required for the formation of only a subcomponent of one complex. The 5' gRNA sequence encompassing the RYAYA and anchor regions and the 3' gRNA oligo(U) tail are both important in complex formation, since their deletion results in a dramatic decrease of some complexes and the absence of others. UV cross-linking experiments identify several proteins which are in contact with gRNA and preedited mRNA in mitochondrial extracts. Proteins of 25 and 90 kDa are highly specific for gRNAs, and the 90-kDa protein binds specifically to gRNA oligo(U) tails. The gRNA-binding proteins exhibit a differential distribution between the four in vitro-formed complexes. These experiments reveal several proteins potentially involved in RNA editing and indicate that multiple recognition elements in gRNAs are used for complex formation.
机译：动素体中的RNA编辑可能采用了高分子复合体，即编辑体，该复合体可能包含指定编辑序列的指导RNA（gRNA）。与gRNA体外形成的特定核糖核蛋白（RNP）复合物（H. U. G？ringer，D。J. Koslowsky，T。H. Morales和K. D. Stuart，Proc。Natl.Acad.Sci。USA，印刷中）是潜在的糖体或其前体。我们发现几个因素对于这些RNP复合物的体外形成很重要，并确定复合物中存在的特定gRNA结合蛋白。在某些条件下，预编辑的mRNA可以促进四种主要的含gRNA的RNP复合物的体外形成，但仅形成一种复合物的一个亚成分是必需的。包含RYAYA和锚定区的5'gRNA序列以及3'gRNA oligo（U）尾部在复合物形成中都很重要，因为它们的缺失会导致某些复合物的急剧减少和其他复合物的缺失。紫外线交联实验确定了与gRNA和线粒体提取物中预编辑的mRNA接触的几种蛋白质。 25 kDa和90 kDa的蛋白质对gRNA具有高度特异性，而90 kDa的蛋白质则与gRNA oligo（U）尾巴特异性结合。 gRNA结合蛋白在四种体外形成的复合物之间表现出差异分布。这些实验揭示了可能参与RNA编辑的几种蛋白质，并表明gRNA中的多种识别元件可用于复合物的形成。

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