Melton D A, Cortese R
Cell. 1979 Dec;18(4):1165-72. doi: 10.1016/0092-8674(79)90229-0.
The transcription of transfer RNA genes (tDNAs) and processing of the transcripts have been studied by injecting cloned tDNAs into Xenopus oocyte nuclei. Three main conclusions can be drawn. First, eucaryotic nuclear tRNA genes, but neither procaryotic nor mitochondrial tRNA genes, are expressed in injected oocytes. While both nematode and yeast tDNAS direct the synthesis of authentic tRNAs, neither E. coli tDNA nor human mitochondrial tDNAs support the synthesis of defined tRNAs when injected into oocytes. Second, competition experiments with co-injected 5S genes and inhibition experiments with alpha-amanitin show that injected tDNAs are transcribed by RNA polymerase III. Third, oocytes injected with a nematode tDNA synthesize a tRNA precursor which is processed post-transcriptionally by removal of a 5' leader sequence. This precursor is found exclusively in the nucleus and is processed in the nucleus before the mature tRNA enters the cytoplasm.
通过将克隆的转运RNA基因(tDNA)注射到非洲爪蟾卵母细胞核中,对转运RNA基因(tDNA)的转录及转录本的加工过程进行了研究。可以得出三个主要结论。第一,真核细胞核tRNA基因,而非原核生物或线粒体tRNA基因,在注射的卵母细胞中得以表达。虽然线虫和酵母的tDNA都能指导合成真正的tRNA,但当将大肠杆菌tDNA或人类线粒体tDNA注射到卵母细胞中时,它们都无法支持合成特定的tRNA。第二,与共注射的5S基因进行的竞争实验以及用α-鹅膏蕈碱进行的抑制实验表明,注射的tDNA由RNA聚合酶III转录。第三,注射了线虫tDNA的卵母细胞合成了一种tRNA前体,该前体在转录后通过去除5'前导序列进行加工。这种前体仅存在于细胞核中,并且在成熟tRNA进入细胞质之前在细胞核中进行加工。