Schulze-Lefert P, Dangl J L, Becker-André M, Hahlbrock K, Schulz W
Department of Biochemistry, Max Planck Institut für Züchtungsforschung, Köln, FRG.
EMBO J. 1989 Mar;8(3):651-6. doi: 10.1002/j.1460-2075.1989.tb03422.x.
We began characterization of the protein--DNA interactions necessary for UV light induced transcriptional activation of the gene encoding chalcone synthase (CHS), a key plant defense enzyme. Three light dependent in vivo footprints appear on a 90 bp stretch of the CHS promoter with a time course correlated with the onset of CHS transcription. We define a minimal light responsive promoter by functional analysis of truncated CHS promoter fusions with a reporter gene in transient expression experiments in parsley protoplasts. Two of the three footprinted sequence 'boxes' reside within the minimal promoter. Replacement of 10 bp within either of these 'boxes' leads to complete loss of light responsiveness. We conclude that these sequences define the necessary cis elements of the minimal CHS promoter's light responsive element. One of the functionally defined 'boxes' is homologous to an element implicated in regulation of genes involved in photosynthesis. These data represent the first example in a plant defense gene of an induced change in protein--DNA contacts necessary for transcriptional activation. Also, our data argue strongly that divergent light induced biosynthetic pathways share common regulatory units.
我们开始对紫外线诱导查尔酮合酶(CHS)基因转录激活所必需的蛋白质 - DNA相互作用进行表征,CHS是一种关键的植物防御酶。在CHS启动子的90 bp片段上出现了三个光依赖性的体内足迹,其时间进程与CHS转录的开始相关。我们通过在欧芹原生质体的瞬时表达实验中对截短的CHS启动子与报告基因的融合体进行功能分析,定义了一个最小光响应启动子。三个足迹序列“框”中的两个位于最小启动子内。替换这两个“框”中任何一个内的10 bp会导致光响应完全丧失。我们得出结论,这些序列定义了最小CHS启动子光响应元件的必需顺式元件。其中一个功能定义明确的“框”与一个参与光合作用相关基因调控的元件同源。这些数据代表了植物防御基因中第一个转录激活所必需的蛋白质 - DNA接触诱导变化的例子。此外,我们的数据有力地表明,不同的光诱导生物合成途径共享共同的调控单元。