Vorst O, Kock P, Lever A, Weterings B, Weisbeek P, Smeekens S
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
Plant J. 1993 Dec;4(6):933-45. doi: 10.1046/j.1365-313x.1993.04060933.x.
Plastocyanin is part of the photosynthetic electron transport chain in the chloroplast and is encoded in the nucleus. Expression of the Arabidopsis thaliana plastocyanin gene is organ specific: high mRNA levels are observed in young green parts of the plant. Furthermore, expression is dependent on the presence of light and functional chloroplasts. When grown in the presence of norflurazon under white light conditions, resulting in the photo-oxidative destruction of the chloroplast, plastocyanin mRNA levels are strongly reduced. A -1579 to -9 promoter fragment confers light-regulated and chloroplast-dependent expression to the beta-glucuronidase reporter gene in transgenic tobacco plants. This suggests that regulation takes place at the level of transcription. A plastocyanin promoter deletion series ranging from -1579 to -121 which was also tested in tobacco, revealed the presence of a strong positive regulating element (PRE) in the -1579 to -705 region. Deletion of this part of the promoter resulted in a approximately 100-fold reduction of GUS expression as measured in mature leaves. Surprisingly, this enhancer-like element was capable of stimulating transcription from a position downstream of its reporter. Moreover, it could also activate a truncated CaMV 35S promoter. Deletion of this element coincides with the loss of chloroplast-dependency of reporter gene expression, as judged by norflurazon treatment of transgenic seedlings. So, the activity of the PRE itself might depend on the presence of functional chloroplasts.
质体蓝素是叶绿体中光合电子传递链的一部分,由细胞核编码。拟南芥质体蓝素基因的表达具有器官特异性:在植物幼嫩的绿色部分观察到较高的mRNA水平。此外,表达依赖于光和功能正常的叶绿体。在白光条件下,当在去甲草酮存在下生长时,叶绿体发生光氧化破坏,质体蓝素mRNA水平会大幅降低。一个-1579至-9的启动子片段赋予转基因烟草植株中β-葡萄糖醛酸酶报告基因光调节和叶绿体依赖性表达。这表明调控发生在转录水平。在烟草中也进行了测试的一个从-1579至-121的质体蓝素启动子缺失系列,揭示了在-1579至-705区域存在一个强正调控元件(PRE)。如在成熟叶片中所测,缺失启动子的这一部分导致GUS表达降低约100倍。令人惊讶的是,这个类似增强子的元件能够从其报告基因下游的一个位置刺激转录。此外,它还能激活一个截短的CaMV 35S启动子。通过对转基因幼苗进行去甲草酮处理判断,缺失这个元件与报告基因表达的叶绿体依赖性丧失同时发生。因此,PRE自身的活性可能依赖于功能正常的叶绿体的存在。