Van Etten R A, Bird J W, Clayton D A
J Biol Chem. 1983 Aug 25;258(16):10104-10.
The 3'-termini of the mitochondrial 12 S and 16 S ribosomal RNAs from mouse L cells have been definitively characterized by mobility-shift RNA sequencing, RNase digestion followed by fingerprinting using two-dimensional homochromatography, and precise mapping of RNA-DNA duplexes using nuclease S1. The results have been correlated with the known DNA sequence of the rRNA region. The vast majority of the 12 S rRNA consists of a family of transcripts whose last template-encoded nucleotide corresponds to a position immediately adjacent to the 5' end of the tRNAVal gene in the DNA sequence. These transcripts are oligoadenylated at their 3'-ends with from 1 to about 5 adenylate residues that are not encoded in the DNA sequence. A minor proportion of the 12 S rRNA ends one nucleotide before the 12 S/tRNAVal gene boundary and is also oligoadenylated. In contrast, the 3'-termini of 16 S rRNA have considerably greater heterogeneity, with the genomic location of the last template-encoded nucleotide varying from the nucleotide immediately adjacent to the 5'-end of the tRNALeuUUR gene to any position up to 7 nucleotides downstream within the tRNALeuUUR gene sequence. These various 16 S rRNA transcripts are oligoadenylated to a somewhat greater degree than the 12 S rRNA. The extent of the 16 S rRNA 3' heterogeneity, as compared to the 12 S rRNA, suggests that the 16 S rRNA 3'-termini may be generated by a mechanism involving termination of transcription rather than by processing of a primary transcript. The data are similar to those reported for human mitochondrial rRNA 3'-termini, and support a general role for adenylation of 3'-termini in the termination or processing of mammalian mitochondrial RNAs.
通过迁移率变动RNA测序、核糖核酸酶消化后使用二维同系层析法进行指纹图谱分析以及使用核酸酶S1对RNA-DNA双链体进行精确作图,已明确鉴定了小鼠L细胞线粒体12S和16S核糖体RNA的3'末端。研究结果已与rRNA区域的已知DNA序列相关联。绝大多数12S rRNA由一族转录本组成,其最后一个模板编码的核苷酸对应于DNA序列中紧邻tRNAVal基因5'端的位置。这些转录本在其3'末端被寡腺苷酸化,带有1至约5个未在DNA序列中编码的腺苷酸残基。一小部分12S rRNA在12S/tRNAVal基因边界前一个核苷酸处结束,并且也被寡腺苷酸化。相比之下,16S rRNA的3'末端具有更大的异质性,最后一个模板编码的核苷酸的基因组位置从紧邻tRNALeuUUR基因5'端的核苷酸到tRNALeuUUR基因序列内下游多达7个核苷酸的任何位置不等。这些不同的16S rRNA转录本比12S rRNA的寡腺苷酸化程度略高。与12S rRNA相比,16S rRNA 3'异质性的程度表明,16S rRNA 3'末端可能是通过涉及转录终止的机制产生的,而不是通过初级转录本的加工产生的。这些数据与报道的人类线粒体rRNA 3'末端的数据相似,并支持3'末端腺苷酸化在哺乳动物线粒体RNA的终止或加工中的一般作用。