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核糖核酸酶P的催化性RNA对tRNA的过度加工。在成熟tRNA序列内对天然tRNA的切割以及底物tRNA构象改变的证据。

Hyperprocessing of tRNA by the catalytic RNA of RNase P. Cleavage of a natural tRNA within the mature tRNA sequence and evidence for an altered conformation of the substrate tRNA.

作者信息

Kikuchi Y, Sasaki N

机构信息

Department of Molecular Biology, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.

出版信息

J Biol Chem. 1992 Jun 15;267(17):11972-6.

PMID:1601867
Abstract

In the transposon copia-related retrovirus-like particles of Drosophila, a 39-nucleotide-long fragment from the 5'-region of Drosophila initiator methionine tRNA (tRNA(iMet)) is used as the primer for copia minus-strand reverse transcription. This primer tRNA(iMet) fragment is thought to be produced by cleavage within the mature tRNA(iMet) sequence. We call this cleavage hyperprocessing. We have previously reported that catalytic RNA of RNase P from Escherichia coli (M1RNA) cleaves the synthetic tRNA(iMet) precursor in vitro at several sites within the mature tRNA sequence. Based on this result, we proposed a model for formation of the primer tRNA fragment involving RNase P. Here we show that natural tRNA(iMet) prepared from Drosophila adult flies can be cleaved by M1RNA. Using mutant tRNA(iMet) substrates, we also show that these cleavages are dependent on the occurrence of an altered conformation of the tRNA substrate. This is evidence that a tRNA can exist in aqueous solution at least in part in an altered conformation.

摘要

在果蝇的转座子类copia逆转录病毒样颗粒中，来自果蝇起始甲硫氨酸tRNA（tRNA(iMet)）5'区域的一段39个核苷酸长的片段被用作copia负链逆转录的引物。这个引物tRNA(iMet)片段被认为是由成熟tRNA(iMet)序列内的切割产生的。我们将这种切割称为超加工。我们之前报道过，来自大肠杆菌的核糖核酸酶P的催化RNA（M1RNA）在体外可在成熟tRNA序列内的几个位点切割合成的tRNA(iMet)前体。基于这一结果，我们提出了一个涉及核糖核酸酶P的引物tRNA片段形成模型。在此我们表明，从果蝇成虫制备的天然tRNA(iMet)可被M1RNA切割。使用突变的tRNA(iMet)底物，我们还表明这些切割依赖于tRNA底物构象改变的发生。这证明了tRNA至少部分可以以改变的构象存在于水溶液中。

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