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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Eukaryotic-type plastid nucleoid protein pTAC3 is essential for transcription by the bacterial-type plastid RNA polymerase
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Eukaryotic-type plastid nucleoid protein pTAC3 is essential for transcription by the bacterial-type plastid RNA polymerase

机译：真核型质体核苷蛋白pTAC3对于细菌型质体RNA聚合酶的转录至关重要

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摘要

Plastid transcription is mediated by two distinct types of RNA pol-ymerases (RNAPs), bacterial-type RNAP (PEP) and phage-type RNAP (NEP). Recent genomic and proteomic studies revealed that higher plants have lost most prokaryotic transcription regulators and have acquired eukaryotic-type proteins during plant evolution. However, in vivo dynamics of chloroplast RNA polymerases and eukaryotic-type plastid nucleoid proteins have not been directly characterized experimentally. Here, we examine the association of the a-subunit of PEP and eukaryotic-type protein, plastid transcriptionally active chromosome 3 (pTAC3) with transcribed regions in vivo by using chloroplast chromatin immunoprecipita-tion (cpChIP) assays. PEP a-subunit preferentially associates with PEP promoters of photosynthesis and rRNA genes, but not with NEP promoter regions, suggesting selective and accurate recognition of PEP promoters by PEP. The cpChIP assays further demonstrate that the peak of PEP association occurs at the promoter-proximal region and declines gradually along the transcribed region. pTAC3 is a putative DNA-binding protein that is localized to chloroplast nucleoids and is essential for PEP-dependent transcription. Density gradient and immunoprecipitation analyses of PEP revealed that pTAC3 is associated with the PEP complex. Interestingly, pTAC3 associates with the PEP complex not only during transcription initiation, but also during elongation and termination. These results suggest that pTAC3 is an essential component of the chloroplast PEP complex. In addition, we demonstrate that light-dependent chloroplast transcription is mediated by light-induced association of the PEP-pTAC3 complex with promoters. This study illustrates unique dynamics of PEP and its associated protein pTAC3 during light-dependent transcription in chloroplasts.
机译：质体转录由两种不同类型的RNA聚合酶（RNAP）介导，即细菌型RNAP（PEP）和噬菌体型RNAP（NEP）。最近的基因组和蛋白质组学研究表明，高等植物在植物进化过程中失去了大多数原核转录调节因子，并获得了真核型蛋白质。但是，叶绿体RNA聚合酶和真核型质体核苷蛋白的体内动力学尚未通过实验直接表征。在这里，我们通过使用叶绿体染色质免疫沉淀（cpChIP）分析检查了PEP和真核生物型蛋白的质子转录活性染色体3（pTAC3）与体内转录区的关联。 PEP a亚基优先与光合作用和rRNA基因的PEP启动子相关，但不与NEP启动子区域相关，这表明PEP对PEP启动子的选择性和准确识别。 cpChIP分析进一步证明，PEP缔合的峰值出现在启动子附近区域，并沿着转录区域逐渐下降。 pTAC3是一种假定的DNA结合蛋白，定位于叶绿体核苷酸，对PEP依赖性转录至关重要。 PEP的密度梯度和免疫沉淀分析表明pTAC3与PEP复合物有关。有趣的是，pTAC3不仅在转录起始过程中，而且在延伸和终止过程中都与PEP复合物缔合。这些结果表明，pTAC3是叶绿体PEP复合物的必需成分。另外，我们证明光依赖性叶绿体转录是由光诱导的PEP-pTAC3复合物与启动子的缔合介导的。这项研究说明了叶绿体中光依赖性转录过程中PEP及其相关蛋白pTAC3的独特动力学。

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    Yusuke Yagi; Yoko lshizaki; Yoichi Nakahira; Yuzuru Tozawa; Takashi Shiina;

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    Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan,lnstitute of Advanced Study, Kyushu University, Fukuoka 812-8581, Japan,Graduate School of Life and Environmental Science, Kyoto Prefectural University, Kyoto 606-8522, Japan;

    Graduate School of Life and Environmental Science, Kyoto Prefectural University, Kyoto 606-8522, Japan;

    Graduate School of Life and Environmental Science, Kyoto Prefectural University, Kyoto 606-8522, Japan;

    Cell-Free Science and Technology Research Center, Ehime University, Matsuyama 790-8577, Japan;

    Graduate School of Life and Environmental Science, Kyoto Prefectural University, Kyoto 606-8522, Japan;

  • 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);
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