INVITRO TRANSCRIPTION OF HUMAN MITOCHONDRIAL-DNA - ACCURATE TERMINATION REQUIRES A REGION OF DNA-SEQUENCE THAT CAN FUNCTION BIDIRECTIONALLY
INVITRO TRANSCRIPTION OF HUMAN MITOCHONDRIAL-DNA - ACCURATE TERMINATION REQUIRES A REGION OF DNA-SEQUENCE THAT CAN FUNCTION BIDIRECTIONALLY
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DOI：
10.1073/pnas.83.17.6277
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发表时间：
1986-09-01
影响因子：
11.1
通讯作者：
CLAYTON, DA
中科院分区：
文献类型：
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作者：
CHRISTIANSON, TW;CLAYTON, DA
哺乳动物线粒体基因组在16S rRNA-tRNALeu基因边界处有一个推定的转录终止位点。我们已经从人类KB细胞中开发了一种体外系统，该系统在该基因边界终止转录。通过S1核酸酶的保护，终止转录本的3”端被定位在16S rRNA-tRNALeu基因边界上游3和4个碱基对，与体内数据一致。通过转录本的高分辨率大小，3”端被映射为7。基因边界下游1个碱基对。在重链启动子或轻链启动子的转录起始处终止具有同等效力。终止所需的所有模板核苷酸序列信息似乎都位于终止位点本身附近。一个意想不到的观察结果是终止区是双向作用的。
Mammalian mitochondrial genomes have a presumptive transcription termination site at the 16S rRNA-tRNALeu gene boundary. We have developed an in vitro system from human KB cells that terminates transcription at this gene boundary. By S1 nuclease protection, the 3'' ends of terminated transcripts were mapped 3 and 4 base pairs upstream of the 16S rRNA-tRNALeu gene boundary, in agreement with in vivo data. By high-resolution sizing of transcripts, the 3'' end was mapped 7 .+-. 1 base pairs downstream from the gene boundary. Termination occurs with equal efficacy from trascriptional initiation at the heavy- or light-strand promoter. All template nucleotide sequence information necessary for termination appears to be located near the termination site itself. An unexpected observation is that the termination region functions bidirectionally.