Template sequences required for transcription of Xenopus laevis mitochondrial DNA from two bidirectional promoters

Template sequences required for transcription of Xenopus laevis mitochondrial DNA from two bidirectional promoters
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从两个双向启动子转录非洲爪蟾线粒体 DNA 所需的模板序列

DOI:
10.1128/mcb.8.7.2917-2924.1988
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发表时间:
1988
影响因子:
5.3
通讯作者:
M. Romanelli
M. Romanelli
中科院分区:
生物学2区
文献类型:
--
作者:
D. Bogenhagen;M. Romanelli

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我们实验室的前期工作表明,非洲爪哇线粒体DNA的转录在体内和体外都是由位于tRNA基因(Phe)和置换环DNA链5‘端之间的双向启动子启动的。与八核苷酸ACGTTATA匹配的共同序列围绕着每个转录起始点。在本研究中,我们使用体外诱变来确定体外特异转录所需的序列。首先,通过缺失突变产生的克隆线粒体DNA模板在体外转录,以确定功能启动子的界限。最详细地研究了位于tRNA(Phe)基因上游约33个核苷酸的双向启动子,称为启动子1。结果证实了这样的假设,即围绕每个起始点的一致的八核苷酸序列是一个重要的启动子元件。合成了编码对称启动子1区的18个碱基对的双链寡核苷酸,并将其克隆到载体中。这种人工合成的寡核苷酸足以支持双向转录。使用该寡核苷酸中的点突变来鉴定共识序列中的关键残基。
Previous work from our laboratory has shown that transcription of Xenopus laevis mitochondrial DNA initiates both in vivo and in vitro from bidirectional promoters located between the gene for tRNA(Phe) and the 5' termini of displacement loop DNA strands. A consensus sequence matching the octanucleotide ACGTTATA surrounds each transcription start site. In the present study, we used in vitro mutagenesis to define sequences required for specific transcription in vitro. First, cloned mitochondrial DNA templates generated by deletion mutagenesis were transcribed in vitro to define the limits of functional promoters. The bidirectional promoter located approximately 33 nucleotides upstream from the gene for tRNA(Phe), termed promoter 1, was studied in greatest detail. The results confirmed the hypothesis that the consensus octanucleotide sequence surrounding each start site is an essential promoter element. A duplex 18-base-pair oligonucleotide encoding the symmetrical promoter 1 region was synthesized and cloned in a plasmid vector. This synthetic oligonucleotide was sufficient to support bidirectional transcription. Point mutations within this oligonucleotide were used to identify critical residues within the consensus sequence.
DOI: --
发表时间: 1986
期刊: The Journal of biological chemistry
影响因子: --
作者:
Cairns,SS;Bogenhagen,DF
通讯作者: Bogenhagen,DF
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DOI: --
发表时间: 1987
期刊: The Journal of biological chemistry
影响因子: --
作者:
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DOI: 10.1126/science.2951850
发表时间: 1987
期刊: Science (New York, N.Y.)
影响因子: --
作者:
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通讯作者: Blattner,FR
DOI: 10.1002/j.1460-2075.1985.tb03817.x
发表时间: 1985-01-01
期刊: EMBO JOURNAL
影响因子: 11.4
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CHANG, DD;HAUSWIRTH, WW;CLAYTON, DA
通讯作者: CLAYTON, DA
DOI: --
发表时间: 1986
期刊: The Journal of biological chemistry
影响因子: --
作者:
Bogenhagen,DF;Yoza,BK;Cairns,SS
通讯作者: Cairns,SS