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一种40千道尔顿的蛋白质特异性结合酵母线粒体mRNA的5'非翻译区。

A 40 kd protein binds specifically to the 5'-untranslated regions of yeast mitochondrial mRNAs.

作者信息

Papadopoulou B, Dekker P, Blom J, Grivell L A

机构信息

Department of Molecular Cell Biology, University of Amsterdam, The Netherlands.

出版信息

EMBO J. 1990 Dec;9(12):4135-43. doi: 10.1002/j.1460-2075.1990.tb07636.x.

DOI:10.1002/j.1460-2075.1990.tb07636.x
PMID:1701144
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC552187/
Abstract

Using a gel mobility shift assay we show that a 40 kd protein (p40), present in extracts of yeast mitochondria, binds specifically to the 5'-untranslated leader of cytochrome c oxidase subunit II mRNA. Binding of p40 to coxII RNA protects an 8-10 nucleotide segment from diethylpyocarbonate modification, indicating that the protein interacts with only a restricted region of the 5'-leader. This segment is located at position -12 with respect to the initiation AUG. Deletion of 10 nucleotides encompassing this site completely abolishes protein binding. Nevertheless, Bal31 deletion analysis within the coxII leader shows that a major part of the leader is essential for p40 binding, suggesting that binding of the protein is also dependent on secondary structural features. p40 binds to other mitochondrial leader mRNAs including those for coxI, coxIII and cyt b. p40 is present in a cytoplasmic (rho0) petite mutant lacking mitochondrial protein synthesis. It is therefore presumably nuclear encoded. The possible biological function of the protein is discussed.

摘要

通过凝胶迁移率变动分析，我们发现酵母线粒体提取物中存在的一种40 kd蛋白（p40）能特异性结合细胞色素c氧化酶亚基II mRNA的5'非翻译前导区。p40与coxII RNA的结合可保护一段8 - 10个核苷酸的片段免受焦碳酸二乙酯修饰，这表明该蛋白仅与5'前导区的一个受限区域相互作用。该片段相对于起始AUG位于 - 12位。缺失包含此位点的10个核苷酸会完全消除蛋白结合。然而，coxII前导区内的Bal31缺失分析表明，前导区的大部分对于p40结合至关重要，这表明该蛋白的结合也依赖于二级结构特征。p40还能结合其他线粒体前导mRNA，包括coxI、coxIII和cyt b的mRNA。p40存在于缺乏线粒体蛋白合成的细胞质（rho0）小菌落突变体中。因此，推测它是由核编码的。本文还讨论了该蛋白可能的生物学功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/18ee3cb2ac8a/emboj00239-0332-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/7271eca44591/emboj00239-0329-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/71ec3e858146/emboj00239-0329-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/8dad7ee02111/emboj00239-0330-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/b5cb774769da/emboj00239-0330-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/fdbc24188ea0/emboj00239-0331-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/9f3e583d9b8f/emboj00239-0332-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/18ee3cb2ac8a/emboj00239-0332-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/7271eca44591/emboj00239-0329-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/71ec3e858146/emboj00239-0329-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/8dad7ee02111/emboj00239-0330-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/b5cb774769da/emboj00239-0330-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/fdbc24188ea0/emboj00239-0331-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/9f3e583d9b8f/emboj00239-0332-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ec0/552187/18ee3cb2ac8a/emboj00239-0332-b.jpg

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Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
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