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RNA Sequence and Base Pairing Effects on Insertion Editing in Trypanosoma brucei

机译：布氏锥虫RNA序列和碱基配对对插入编辑的影响

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摘要

RNA editing inserts and deletes uridylates (U's) in kinetoplastid mitochondrial pre-mRNAs by a series of enzymatic steps. Small guide RNAs (gRNAs) specify the edited sequence. Editing, though sometimes extensive, is precise. The effects of mutating pre-mRNA and gRNA sequences in, around, and upstream of the editing site on the specificity and efficiency of in vitro insertion editing were examined. U's could be added opposite guiding pyrimidines, but guiding purines, particularly A's, were required for efficient ligation. A base pair between mRNA and gRNA immediately upstream of the editing site was not required for insertion editing, although it greatly enhanced its efficiency and accuracy. In addition, a gRNA/mRNA duplex upstream of the editing site enhanced insertion editing when it was close to the editing site, but prevented cleavage, and hence editing, when immediately adjacent to the editing site. Thus, several aspects of mRNA-gRNA interaction, as well as gRNA base pairing with added U's, optimize editing efficiency, although they are not required for insertion editing.
机译：RNA编辑通过一系列酶促步骤在动质体线粒体前mRNA中插入和删除尿酸（U's）。小指南RNA（gRNA）指定编辑的序列。编辑虽然有时很广泛，但却很精确。研究了在编辑位点附近，周围和上游突变前mRNA和gRNA序列对体外插入编辑的特异性和效率的影响。可以在导向嘧啶的对面添加U's，但有效连接需要导向嘌呤，特别是A'。插入编辑不需要直接位于编辑位点上游的mRNA和gRNA之间的碱基对，尽管它大大提高了效率和准确性。另外，当gRNA / mRNA双链体靠近编辑位点时，其上游的gRNA / mRNA双链体增强了插入编辑，但是当其紧邻编辑位点时阻止了切割，从而防止了编辑。因此，mRNA-gRNA相互作用的几个方面以及与添加的U配对的gRNA碱基配对可优化编辑效率，尽管插入编辑并不需要它们。

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