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首页> 美国卫生研究院文献>Nucleic Acids Research >Allosteric regulation of tRNA import: interactions between tRNA domains at the inner membrane of Leishmania mitochondria
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Allosteric regulation of tRNA import: interactions between tRNA domains at the inner membrane of Leishmania mitochondria

机译：tRNA导入的变构调节：利什曼原虫线粒体内膜上的tRNA结构域之间的相互作用

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摘要

Import of nucleus-encoded tRNAs into the mitochondria of the kinetoplastid protozoon Leishmania involves recognition of specific import signals by the membrane-bound import machinery. Multiple signals on different tRNA domains may be present, and further, importable RNAs interact positively (Type I) or negatively (Type II) with one another at the inner membrane in vitro. By co-transfection assays, it is shown here that tRNATyr (Type I) transiently stimulates the rate of entry of tRNAIle (Type II) into Leishmania mitochondria in transfected cells, and conversely, is inhibited by tRNAIle. Truncation and mutagenesis experiments led to the co-localization of the effector and import activities of tRNATyr to the D domain, and those of tRNAIle to the variable region–T domain (V-T region), indicating that both activities originate from a single RNA–receptor interaction. A third tRNA, human tRNALys, is imported into Leishmania mitochondria in vitro as well as in vivo. This tRNA has Type I and Type II motifs in the D domain and the V-T region, respectively, and shows both Type I and Type II effector activities. Such dual-type tRNAs may interact simultaneously with the Type I and Type II binding sites of the inner membrane import machinery.
机译：将核编码的tRNA导入到运动质体原生动物利什曼原虫的线粒体中，需要通过膜结合的导入机制识别特定的导入信号。可能存在不同tRNA结构域上的多个信号，此外，可导入的RNA在体外在内膜上彼此发生正向相互作用（I型）或负向相互作用（II型）。通过共转染测定法，这里显示了tRNA Tyr （I型）瞬时刺激了转染细胞中tRNA Ile （II型）进入利什曼原虫线粒体的速率。反之，则被tRNA Ile 抑制。截短和诱变实验导致tRNA Tyr 的效应子和导入活性共定位于D结构域，而tRNA Ile 的效应子和导入活性共定位于可变区–T结构域（VT区域），表明这两种活性都源自单个RNA-受体相互作用。第三种tRNA，即人tRNA Lys ，可在体内和体外导入利什曼原虫线粒体中。该tRNA分别在D结构域和V-T区域具有I型和II型基序，并显示I型和II型效应子活性。这样的双型tRNA可以与内膜导入机制的I型和II型结合位点同时相互作用。

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