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COP1的N端片段的表达对拟南芥中光调节的幼苗发育具有显性负效应。

Expression of an N-terminal fragment of COP1 confers a dominant-negative effect on light-regulated seedling development in Arabidopsis.

作者信息

McNellis T W, Torii K U, Deng X W

机构信息

Department of Biology, Yale University, New Haven, Connecticut 06520-8104, USA.

出版信息

Plant Cell. 1996 Sep;8(9):1491-503. doi: 10.1105/tpc.8.9.1491.

Abstract

CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) is an essential regulatory gene that plays a role in light control of seedling development in Arabidopsis. The COP1 protein possesses three recognizable structural domains: a RING finger zinc binding domain near the N terminus, followed by a coiled-coll domain and a domain with WD-40 repeats in the C-terminal half. To determine whether COP1 acts specifically as a light-inactivable repressor of photomorphogenic development and to elucidate the functional roles of the specific structural domains, mutant cDNAs encoding the N-terminal 282 amino acids (N282) of COP1 were expressed and analyzed in transgenic plants. High-level expression of the N282 fragment caused a dominant-negative phenotype similar to that of the loss-of-function cop1 mutants. The phenotypic characteristics include hypersensitivity of hypocotyl elongation to inhibition by white, blue, red, and far-red light stimuli. In the dark, N282 expression led to pleiotropic photomorphogenic cotyledon development, including cellular differentiation, plastid development, and gene expression, although it has no significant effect on the hypocotyl elongation. However, N282 expression had a minimal effect on the expression of stress- and pathogen-inducible genes. These observations support the hypothesis that COP1 is directly involved in the light control of seedling development and that it acts as a repressor of photomorphogenesis. Further, the results imply that the N282 COP1 fragment, which contains the zinc binding and colled-coil domains, is capable of interacting with either downstream targets or with the endogenous wild-type COP1, thus interfering with normal regulatory processes. The fact the N282 is able to interact with N282 and full-length COP1 in yeast provided evidence for the latter possibility.

摘要

组成型光形态建成1(COP1)是一个重要的调控基因,在拟南芥幼苗发育的光控制中发挥作用。COP1蛋白具有三个可识别的结构域:靠近N端的一个环指锌结合结构域,接着是一个卷曲螺旋结构域和在C端后半部分的一个具有WD-40重复序列的结构域。为了确定COP1是否专门作为光形态建成发育的光失活阻遏物,并阐明特定结构域的功能作用,编码COP1 N端282个氨基酸(N282)的突变cDNA在转基因植物中进行了表达和分析。N282片段的高水平表达导致了与功能缺失型cop1突变体相似的显性负性表型。表型特征包括下胚轴伸长对白光、蓝光、红光和远红光刺激抑制的超敏反应。在黑暗中,N282的表达导致了多效性的光形态建成子叶发育,包括细胞分化、质体发育和基因表达,尽管它对下胚轴伸长没有显著影响。然而,N282的表达对胁迫和病原体诱导基因的表达影响最小。这些观察结果支持了COP1直接参与幼苗发育的光控制且作为光形态建成阻遏物的假说。此外,结果表明包含锌结合和卷曲螺旋结构域的N282 COP1片段能够与下游靶标或内源性野生型COP1相互作用,从而干扰正常的调控过程。N282能够在酵母中与N282和全长COP1相互作用这一事实为后一种可能性提供了证据。

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