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在体外系统中，从芥菜（Sinapis alba L.）的质体中准确转录一个编码蛋白质的质体基因所需的 DNA 序列要求。

DNA sequence requirements for the accurate transcription of a protein-coding plastid gene in a plastid in vitro system from mustard (Sinapis alba L.).

机构信息

Biologisches Institut II, Universität Freiburg, Schänzlestr. 1, D-7800 Freiburg, FRG.

出版信息

EMBO J. 1984 Aug;3(8):1697-704. doi: 10.1002/j.1460-2075.1984.tb02034.x.

DOI:10.1002/j.1460-2075.1984.tb02034.x
PMID:16453540
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC557584/
Abstract

A nuclease-treated plastid extract from mustard (Sinapis alba L.) allows efficient transcription of cloned plastid DNA templates. In this in vitro system, the major runoff transcript of the truncated gene for the 32 000 mol. wt. photosystem II protein was accurately initiated from a site close to or identical with the in vivo start site. By using plasmids with deletions in the 5'-flanking region of this gene as templates, a DNA region required for efficient and selective initiation was detected 28-35 nucleotides upstream of the transcription start site. This region contains the sequence element TTGACA, which matches the consensus sequence for prokaryotic ;-35' promoter elements. In the absence of this region, a region 13-27 nucleotides upstream of the start site still enables a basic level of specific transcription. This second region contains the sequence element TATATAA, which matches the consensus sequence for the ;TATA' box of genes transcribed by RNA polymerase II (or B). The region between the ;TATA'-like element and the transcription start site is not sufficient but may be required for specific transcription of the plastid gene. This latter region contains the sequence element TATACT, which resembles the prokaryotic ;-10' (Pribnow) box. Based on the structural and transcriptional features of the 5' upstream region, a ;promoter switch' mechanism is proposed, which may account for the developmentally regulated expression of this plastid gene.

摘要

从芥菜（Sinapis alba L.）中经核酸酶处理的质体提取物允许对克隆的质体 DNA 模板进行有效的转录。在这个体外系统中，截断的 32000 分子量光系统 II 蛋白基因的主要流路转录本从靠近或与体内起始位点相同的位点准确起始。通过使用该基因 5'-侧翼区缺失的质粒作为模板，检测到一个在转录起始位点上游 28-35 个核苷酸的 DNA 区域，该区域对于有效和选择性起始是必需的。该区域包含 TTGACA 序列元件，与原核生物的 -35'启动子元件的共有序列相匹配。在没有该区域的情况下，起始位点上游 13-27 个核苷酸的区域仍然能够实现基本水平的特异性转录。第二个区域包含 TATATAA 序列元件，与由 RNA 聚合酶 II（或 B）转录的基因的“TATA”框的共有序列相匹配。位于“TATA”样元件和转录起始位点之间的区域不是必需的，但可能是质体基因特异性转录所必需的。该后一区域包含 TATACT 序列元件，类似于原核生物的-10'（Pribnow）盒。基于 5'上游区域的结构和转录特征，提出了一种“启动子开关”机制，该机制可能解释了该质体基因的发育调控表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/2cff3e9a2d14/emboj00312-0038-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/1f940d0137cd/emboj00312-0036-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/933048f89f51/emboj00312-0036-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/0eb75e537350/emboj00312-0037-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/8da8e7a37cc3/emboj00312-0037-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/2cff3e9a2d14/emboj00312-0038-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/1f940d0137cd/emboj00312-0036-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/933048f89f51/emboj00312-0036-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/0eb75e537350/emboj00312-0037-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/8da8e7a37cc3/emboj00312-0037-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc5/557584/2cff3e9a2d14/emboj00312-0038-a.jpg

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本文引用的文献

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Characterization of the 32,000 Dalton Chloroplast Membrane Protein: III. Probing Its Biological Function in Spirodela.叶绿体膜蛋白 32000 道尔顿的特性研究：III. 探讨其在满江红中的生物学功能。
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Nucleotide sequence of the gene for the M(r) 32,000 thylakoid membrane protein from Spinacia oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of M(r) 38,950.
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