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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Arabidopsis circadian clock protein, TOC1, is a DNA-binding transcription factor
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Arabidopsis circadian clock protein, TOC1, is a DNA-binding transcription factor

机译：拟南芥生物钟蛋白TOC1是DNA结合转录因子

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摘要

The first described feedback loop of the Arabidopsis circadian clock is based on reciprocal regulation between TIMING OF CAB EXPRESSION 1 (TOC1) and CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1)/LATE ELONGATED HYPOCOTYL (LHY). CCA1 and LHY are Myb transcription factors that bind directly to the TOC1 promoter to negatively regulate its expression. Conversely, the activity of TOC1 has remained less well characterized. Genetic data support that TOC1 is necessary for the reactivation of CCA1/LHY, but there is little description of its biochemical function. Here we show that TOC1 occupies specific genomic regions in the CCA 1 and LHY promoters. Purified TOC1 binds directly to DNA through its CCT domain, which is similar to known DNA-binding domains. Chemical induction and transient overexpression of TOC1 in Arabidopsis seedlings cause repression of CCA1/LHY expression, demonstrating that TOC1 can repress direct targets, and mutation or deletion of the CCT domain prevents this repression showing that DNA-binding is necessary for TOC1 action. Furthermore, we use the Gal4/UAS system in Arabidopsis to show that TOC1 acts as a general transcriptional repressor, and that repression activity is in the pseudoreceiver domain of the protein. To identify the genes regulated by TOC1 on a genomic scale, we couple TOC1 chemical induction with microarray analysis and identify previously unexplored potential TOC1 targets and output pathways. Taken together, these results define a biochemical action for the core clock protein TOC1 and refine our perspective on how plant clocks function.
机译：拟南芥生物钟的第一个描述的反馈回路基于CAB表达1（TOC1）的时序与CIRCADIAN时钟关联1（CCA1）/晚期加长的双氢代叶酸（LHY）之间的相互调节。 CCA1和LHY是Myb转录因子，直接与TOC1启动子结合，从而负面调节其表达。相反，TOC1的活性仍然不太好表征。遗传数据支持TOC1对于CCA1 / LHY的重新激活是必需的，但对其生化功能的描述很少。在这里，我们显示TOC1在CCA 1和LHY启动子中占据特定的基因组区域。纯化的TOC1通过其CCT域直接与DNA结合，这与已知的DNA结合域相似。拟南芥幼苗中TOC1的化学诱导和瞬时过表达引起CCA1 / LHY表达的抑制，表明TOC1可以抑制直接靶标，CCT域的突变或缺失阻止了这种抑制，表明DNA结合对于TOC1的作用是必需的。此外，我们在拟南芥中使用Gal4 / UAS系统来显示TOC1充当一般的转录阻遏物，并且阻遏活性在蛋白质的伪受体结构域中。为了在基因组规模上鉴定由TOC1调控的基因，我们将TOC1化学诱导与微阵列分析相结合，并鉴定以前未开发的潜在TOC1靶标和输出途径。综上所述，这些结果定义了核心时钟蛋白TOC1的生化作用，并完善了我们对植物时钟功能的看法。

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    Joshua M. Gendron; Jose L. Pruneda-Paz; Colleen J. Doherty; Andrew M. Gross; S. Earl Kang; Steve A. Kay;

  • 作者单位

    Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, CA 92093-0130;

    Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, CA 92093-0130;

    Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, CA 92093-0130;

    Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, CA 92093-0130;

    Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, CA 92093-0130;

    Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, CA 92093-0130;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    transcriptional regulation; transcriptional potential; gene networks circadian outputs; diurnal;

    机译：转录调控;转录潜能基因网络的昼夜节律输出;日间;

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