Anderson S L, Teakle G R, Martino-Catt S J, Kay S A
NSF Center for Biological Timing, University of Virginia, Charlottesville 22903.
Plant J. 1994 Oct;6(4):457-70. doi: 10.1046/j.1365-313x.1994.6040457.x.
The Arabidopsis CAB2 promoter was used to define the terminal genomic targets that are subject to regulation by the circadian clock. An in vivo cab::luciferase bioluminescent marker was used to enable the assaying of the expression of chimeric constructs with unprecedented sensitivity and time resolution in living seedlings. Dissection of -322 to +1 of the CAB2 promoter has revealed several interesting features: it was demonstrated that the 323 bp fragment contains at least one strong general positive element. The positive element contains an ACGT core sequence specifically bound by a protein activity, termed CUF-1, and contributes to high level expression but is not required for phytochrome- or circadian-regulation. Moreover, a 78 bp domain was defined that confers both circadian- and phytochrome-regulation upon heterologous promoters. Conserved GATA sequences within the 78 bp regulatory domain are specifically bound by a protein factor designated CGF-1. The binding specificity of CGF-1 appears to be related to the GT-family of trihelix DNA-binding proteins. The role of these DNA-protein interactions is discussed in terms of clock- and phytochrome regulation, and their relevance as targets for pathways defined by photomorphogenic mutants.
拟南芥CAB2启动子用于确定受生物钟调控的末端基因组靶点。体内cab::荧光素酶生物发光标记用于在活体幼苗中以前所未有的灵敏度和时间分辨率检测嵌合构建体的表达。对CAB2启动子-322至+1区域的剖析揭示了几个有趣的特征:已证明323 bp片段包含至少一个强的通用正向元件。该正向元件包含一个由一种称为CUF-1的蛋白质活性特异性结合的ACGT核心序列,有助于高水平表达,但不是光敏色素或生物钟调节所必需的。此外,定义了一个78 bp的结构域,它赋予异源启动子生物钟和光敏色素调节功能。78 bp调节结构域内保守的GATA序列被一种称为CGF-1的蛋白质因子特异性结合。CGF-1的结合特异性似乎与三螺旋DNA结合蛋白的GT家族有关。从生物钟和光敏色素调节方面讨论了这些DNA-蛋白质相互作用的作用,以及它们作为光形态建成突变体所定义途径靶点的相关性。