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首页> 外文期刊>Molecular and Cellular Biology >Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene.
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Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene.

机译：表征组成性活性水稻肌动蛋白基因启动子转录的顺式作用元件的表征。

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The promoter of the constitutively expressed rice (Oryza sativa) actin 1 gene (Act1) is highly active in transformed rice plants (W. Zhang, D. McElroy, and R. Wu, Plant Cell 3:1150-1160, 1991). A region 834 bp upstream of the Act1 transcription initiation site contains all the regulatory elements necessary for maximal gene expression in transformed rice protoplasts (D. McElroy, W. Zhang, J. Cao, and R. Wu, Plant Cell 2:163-171, 1990). We have constructed a series of Act1 promoter deletions fused to a bacterial beta-glucuronidase reporter sequence (Gus). Transient expression assays in transformed rice protoplasts, as well as transformed maize cells and tissues, identified two distinct cis-acting regulatory elements in the Act1 promoter. A 38-bp poly(dA-dT) region was found to be a positive regulator of Act1 promoter activity. Deletion of the poly(dA-dT) element lowered Gus expression by at least threefold compared with expression produced by the full-length Act1 promoter. By gel retardation and footprinting, we identified a ubiquitous rice protein which specifically recognizes this poly(dA-dT) element in the constitutively active Act1 promoter. A CCCAA pentamer repeat-containing region was found to be a negative regulator of the Act1 promoter in transformed rice protoplasts. Transient expression assays in different maize cells and tissues with use of the Act1 deletion constructs suggested that the CCCAA pentamer repeat region functions in a complex tissue-specific manner. A CCCAA-binding protein was detected only in root extracts.
机译：组成型表达的水稻（Oryza sativa）肌动蛋白1基因（Act1）的启动子在转化的水稻植物中具有很高的活性（W.Zhang，D.McElroy和R.Wu，Plant Cell 3：1150-1160，1991）。 Act1转录起始位点上游834 bp的区域包含在转化的水稻原生质体中最大基因表达所需的所有调控元件（D. McElroy，W。Zhang，J。Cao和R. Wu，植物细胞2：163-171 （1990年）。我们已经构建了一系列与细菌β-葡萄糖醛酸苷酶报道序列（Gus）融合的Act1启动子缺失。转化的水稻原生质体以及转化的玉米细胞和组织中的瞬时表达测定确定了Act1启动子中两个不同的顺式作用调控元件。发现一个38 bp的poly（dA-dT）区是Act1启动子活性的正调节剂。与全长Act1启动子产生的表达相比，poly（dA-dT）元件的缺失使Gus表达降低了至少三倍。通过凝胶阻滞和足迹，我们发现了一种普遍存在的大米蛋白，它可以特异性识别组成性活性Act1启动子中的这种poly（dA-dT）元素。发现含有CCCAA五聚体重复序列的区域是转化的水稻原生质体中Act1启动子的负调控子。使用Act1缺失构建体在不同玉米细胞和组织中进行的瞬时表达测定表明，CCCAA五聚体重复区以复杂的组织特异性方式起作用。仅在根提取物中检测到CCCAA结合蛋白。

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