Analysis of the Alternative Oxidase Promoters from Soybean1
Analysis of the Alternative Oxidase Promoters from Soybean1
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DOI：
10.1104/pp.103.028183
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发表时间：
2003-11
期刊：
影响因子：
7.4
通讯作者：
David Thirkettle-Watts;T. McCabe;R. Clifton;C. Moore;P. Finnegan;D. Day;J. Whelan
中科院分区：
文献类型：
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作者：
David Thirkettle-Watts;T. McCabe;R. Clifton;C. Moore;P. Finnegan;D. Day;J. Whelan
交替氧化酶（Aox）是一种核编码的线粒体蛋白。在大豆（Glycine max）中，该基因家族的三个成员已被证明在正常植物发育期间和对胁迫的响应中差异表达。为了检测Aox启动子的功能，获得了所有三个大豆基因的基因组片段：Aox 1、Aox 2a和Aox 2b。在大豆悬浮培养细胞的瞬时转化和拟南芥的稳定转化期间，这些片段的紧邻编码区上游的区域用于驱动β-葡萄糖醛酸酶（GUS）表达。GUS报告基因在大豆细胞中的表达模式与免疫印迹法测定的各种内源Aox蛋白的存在或不存在一致。上游区域的不同部分的缺失鉴定了负责大豆细胞中Aox基因表达的正调控和负调控的序列。在拟南芥植物中的报告基因分析表明，差异组织表达模式驱动的三个上游区域，类似于大豆中的内源蛋白质的报告。还检测了拟南芥Aox基因家族的所有五个成员的表达谱，以与大豆上游片段驱动的GUS表达进行比较。尽管来自大豆Aox 2a和Aox 2b的上游片段的启动子活性在拟南芥中显示出与在大豆中相同的组织特异性，但在拟南芥的所有组织中最显著表达的内源基因是Aox 1型。因此，尽管Aox表达的调节通常似乎涉及不同物种中的相同信号，但Aox的不同直系同源物可能对这些信号有不同的响应。大豆Aox基因和相似表达的拟南芥Aox基因之间的上游序列的比较确定了共同的基序。
Alternative oxidase (Aox) is a nuclear-encoded mitochondrial protein. In soybean (Glycine max), the three members of the gene family have been shown to be differentially expressed during normal plant development and in response to stresses. To examine the function of the Aox promoters, genomic fragments were obtained for all three soybean genes: Aox1, Aox2a, and Aox2b. The regions of these fragments immediately upstream of the coding regions were used to drive β-glucuronidase (GUS) expression during transient transformation of soybean suspension culture cells and stable transformation of Arabidopsis. The expression patterns of the GUS reporter genes in soybean cells were in agreement with the presence or absence of the various endogenous Aox proteins, determined by immunoblotting. Deletion of different portions of the upstream regions identified sequences responsible for both positive and negative regulation of Aox gene expression in soybean cells. Reporter gene analysis in Arabidopsis plants showed differential tissue expression patterns driven by the three upstream regions, similar to those reported for the endogenous proteins in soybean. The expression profiles of all five members of the Arabidopsis Aox gene family were examined also, to compare with GUS expression driven by the soybean upstream fragments. Even though the promoter activity of the upstream fragments from soybean Aox2a and Aox2b displayed the same tissue specificity in Arabidopsis as they do in soybean, the most prominently expressed endogenous genes in all tissues of Arabidopsis were of the Aox1 type. Thus although regulation of Aox expression generally appears to involve the same signals in different species, different orthologs of Aox may respond variously to these signals. A comparison of upstream sequences between soybean Aox genes and similarly expressed Arabidopsis Aox genes identified common motifs.