Giraud Estelle, Van Aken Olivier, Ho Lois H M, Whelan James
Australian Research Council Centre of Excellence in Plant Energy Biology, University of Western Australia, Crawley 6009, Western Australia, Australia.
Plant Physiol. 2009 Jul;150(3):1286-96. doi: 10.1104/pp.109.139782. Epub 2009 May 29.
Plant cells integrate signals from external sources and from organelles to regulate gene expression, referred to as anterograde and retrograde signaling, respectively. Functional characterization of the promoter of ALTERNATIVE OXIDASE1a (AOX1a) from Arabidopsis (Arabidopsis thaliana), a marker for mitochondrial retrograde response, was carried out by testing the ability of the AOX1a promoter to drive expression of the reporter gene GUS. This approach identified a strong repressor element, designated the B element, that was necessary for an increased promoter activity in response to the mitochondrial complex I inhibitor rotenone. This element overlaps with a previously identified potential binding site for the transcription factor ABSCISIC ACID INSENSITIVE4 (ABI4). AOX1a promoter activity was fully derepressed in abi4 mutants and was unresponsive to rotenone. Furthermore, deletion of the B element of the AOX1a promoter resulted in increased GUS staining activity compared to the wild-type promoter in transgenic plants. Binding of the ABI4 transcription factor to this region of the AOX1a promoter was demonstrated by electromobility shift and yeast one-hybrid assays. Analysis of transcript abundance for AOX1a in abi4 mutant lines revealed significantly increased levels of AOX1a mRNA that could not be further induced by rotenone, consistent with the role of ABI4 as a repressor that is derepressed in response to rotenone. These results show that ABI4 plays a central role in mediating mitochondrial retrograde signals to induce the expression of AOX1a. Furthermore, they provide a molecular link between mitochondrial and chloroplast retrograde signaling, as ABI4 has been previously shown to act downstream of at least two chloroplast retrograde signaling pathways.
植物细胞整合来自外部来源和细胞器的信号以调节基因表达,分别称为正向和逆向信号传导。通过测试交替氧化酶1a(AOX1a)启动子驱动报告基因GUS表达的能力,对拟南芥(Arabidopsis thaliana)中线粒体逆向反应的标志物AOX1a启动子进行了功能表征。该方法鉴定出一个强抑制元件,命名为B元件,它是响应线粒体复合体I抑制剂鱼藤酮而增加启动子活性所必需的。该元件与先前鉴定的转录因子脱落酸不敏感4(ABI4)的潜在结合位点重叠。AOX1a启动子活性在abi4突变体中完全去抑制,并且对鱼藤酮无反应。此外,与转基因植物中的野生型启动子相比,AOX1a启动子B元件的缺失导致GUS染色活性增加。通过电泳迁移率变动分析和酵母单杂交试验证明了ABI4转录因子与AOX1a启动子的该区域结合。对abi4突变体系中AOX1a转录丰度的分析显示,AOX1a mRNA水平显著增加,且不能被鱼藤酮进一步诱导,这与ABI4作为响应鱼藤酮而去抑制的阻遏物的作用一致。这些结果表明,ABI4在介导线粒体逆向信号以诱导AOX1a表达中起核心作用。此外,它们提供了线粒体和叶绿体逆向信号传导之间的分子联系,因为先前已表明ABI4在至少两条叶绿体逆向信号传导途径的下游起作用。