Timko M P, Kausch A P, Castresana C, Fassler J, Herrera-Estrella L, Van den Broeck G, Van Montagu M, Schell J, Cashmore A R
Nature. 1985;318(6046):579-82. doi: 10.1038/318579a0.
Light regulates many varied physiological and developmental phenomena during plant growth and differentiation, including the formation of a photosynthetically competent chloroplast from a proplastid. The expression of ribulose 1,5-bisphosphate carboxylase small subunit (rbcS) genes is regulated by light in a development- and tissue-specific manner2,3. In some plant species, phytochrome has been demonstrated to mediate this response, and photoregulation of rbcS expression occurs at least in part at the level of transcription. We have shown previously that a 5'-noncoding fragment (4-973 base pairs (bp) upstream of the messenger RNA cap site) of the pea rbcS ss3.6 gene contains all of the nucleotide sequence information necessary to direct the photoregulated expression of a bacterial chloramphenicol acetyltransferase (cat) gene in tobacco. Consistent with these findings, Morelli et al.11 have shown by deletion analysis of a second rbcS gene promoter, that the sequences required for photoregulated expression of rbcS E9 reside within the 5'-noncoding region. They identified an upstream region of approximately 700 bp needed for maximum transcription but not light-dark regulation, and a region from -35 to -2 bp which included the TATA box and contained the necessary information for light responsiveness. We now demonstrate that regulatory sequences 5' distal to the rbcS ss3.6 TATA box and transcriptional start site not only contain the information necessary for maximum expression, but also confer photoregulation. These upstream regulatory sequences function independently of orientation when fused to their homologous promoter or a heterologous promoter.
在植物生长和分化过程中,光调节着许多不同的生理和发育现象,包括从原质体形成具有光合能力的叶绿体。核酮糖-1,5-二磷酸羧化酶小亚基(rbcS)基因的表达受光以发育和组织特异性方式调节2,3。在一些植物物种中,已证明光敏色素介导这种反应,并且rbcS表达的光调节至少部分发生在转录水平。我们先前已经表明,豌豆rbcS ss3.6基因的一个5'-非编码片段(信使RNA帽位点上游4-973个碱基对(bp))包含指导烟草中细菌氯霉素乙酰转移酶(cat)基因光调节表达所需的所有核苷酸序列信息。与这些发现一致,莫雷利等人11通过对第二个rbcS基因启动子的缺失分析表明,rbcS E9光调节表达所需的序列位于5'-非编码区内。他们确定了一个约700 bp的上游区域,该区域是最大转录所必需的,但不是光暗调节所必需的,以及一个从-35到-2 bp的区域,该区域包括TATA框并包含光反应性所需的信息。我们现在证明,rbcS ss3.6 TATA框和转录起始位点5'端的调控序列不仅包含最大表达所需的信息,还赋予光调节作用。当与它们的同源启动子或异源启动子融合时,这些上游调控序列的功能与方向无关。