Hopper A K, Kurjan J
Nucleic Acids Res. 1981 Feb 25;9(4):1019-29. doi: 10.1093/nar/9.4.1019.
In vivo yeast precursor tRNAs have been identified using a modification of the Northern-hybridization procedure. Two species of pre-tRNA Tyr, 1 species of pre-tRNA Ser2 and 2 species of pre-tRNA Serminor have been found in all yeast strains examined, including parental strains and strains harboring mutations affecting tRNA function. One of the tRNA Tyr strains harboring and one of the pre-tRNA SerUCG are the same size as the unspliced pre-tRNAs which accumulate in the yeast mutant rna1. The in vivo tRNA Tyr precursors detected in these studies also appear similar with the RNA species identified when cloned yeast tRNA Tyr is transcribed and processed by Xenopus oocytes and/or Xenopus extracts. We have also studied the precursor and mature tRNA Tyr species from 22 mutants which contain mutations in the SUP4 tyrosine-inserting suppressor locus. The RNA from 2 mutants mapping at the G52 position showing an aberrantly migrating "mature" tRNA Tyr. Although several of those cloned mutant genes showed transcript products of altered size in in vitro transcription studies (1), we did not detect such altered transcripts in vivo.
通过对Northern杂交程序进行改进，已在体内鉴定出酵母前体tRNA。在所有检测的酵母菌株中，包括亲本菌株和携带影响tRNA功能突变的菌株，发现了两种前体tRNA Tyr、一种前体tRNA Ser2和两种前体tRNA Serminor。其中一种携带tRNA Tyr的菌株和一种前体tRNA SerUCG的大小与在酵母突变体rna1中积累的未剪接前体tRNA相同。在这些研究中检测到的体内tRNA Tyr前体也与通过非洲爪蟾卵母细胞和/或非洲爪蟾提取物转录和加工克隆的酵母tRNA Tyr时鉴定出的RNA种类相似。我们还研究了来自22个在SUP4酪氨酸插入抑制基因座中含有突变的突变体的前体和成熟tRNA Tyr种类。来自定位在G52位置的2个突变体的RNA显示出异常迁移的“成熟”tRNA Tyr。尽管在体外转录研究中，其中几个克隆的突变基因显示出大小改变的转录产物(1)，但我们在体内未检测到这种改变的转录本。
