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人类线粒体DNA的体外转录：精确终止需要一个能够双向发挥作用的DNA序列区域。

In vitro transcription of human mitochondrial DNA: accurate termination requires a region of DNA sequence that can function bidirectionally.

作者信息

Christianson T W, Clayton D A

出版信息

Proc Natl Acad Sci U S A. 1986 Sep;83(17):6277-81. doi: 10.1073/pnas.83.17.6277.

DOI:10.1073/pnas.83.17.6277
PMID:3018722
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC386486/
Abstract

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 transcriptional 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.

摘要

哺乳动物线粒体基因组在16S rRNA - tRNALeu基因边界处有一个假定的转录终止位点。我们从人KB细胞中开发了一种体外系统，该系统在此基因边界处终止转录。通过S1核酸酶保护实验，终止转录本的3'端被定位在16S rRNA - tRNALeu基因边界上游3和4个碱基对处，这与体内数据一致。通过对转录本进行高分辨率大小测定，3'端被定位在基因边界下游7±1个碱基对处。从重链或轻链启动子开始转录，终止的效率相同。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。一个意外的发现是终止区域具有双向功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/030734be94f0/pnas00321-0056-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/d4bbaf4d3bb7/pnas00321-0054-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/4319e118e0e6/pnas00321-0055-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/a2b0030ed4b3/pnas00321-0055-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/030734be94f0/pnas00321-0056-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/d4bbaf4d3bb7/pnas00321-0054-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/4319e118e0e6/pnas00321-0055-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/a2b0030ed4b3/pnas00321-0055-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fdf/386486/030734be94f0/pnas00321-0056-a.jpg

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引用本文的文献

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Initiation of mtDNA transcription is followed by pausing, and diverges across human cell types and during evolution.线粒体DNA转录起始后会出现暂停现象，并且在不同人类细胞类型以及进化过程中存在差异。
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Two RNAs or DNAs May Artificially Fuse Together at a Short Homologous Sequence (SHS) during Reverse Transcription or Polymerase Chain Reactions, and Thus Reporting an SHS-Containing Chimeric RNA Requires Extra Caution.

本文引用的文献

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7
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Identification of the 3'-ends of the two mouse mitochondrial ribosomal RNAs. The 3'-end of 16 S ribosomal RNA contains nucleotides encoded by the gene for transfer RNALeuUUR.两种小鼠线粒体核糖体RNA 3'端的鉴定。16S核糖体RNA的3'端含有由亮氨酸转运RNA基因（UUR）编码的核苷酸。
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