Dynamism of plastid gene expression by intracellular communication

细胞内通讯的质体基因表达动态

• 批准号: 14340252
• 负责人: SUGITA Mamoru
• 金额: $ 10.5万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14340252/
• 关键词: Chloroplast; Physcommitrella patens; Plastid transformation; Relic gene; arginine tRNA gene; Plastid RNA polymerase; plastid sigma factor; PPR protein; RNA編集; タバコ; DNAチップ; 概日リズム; Lhcb遺伝子; ミトコンドリア; 翻訳開始コドン; シグマ因子; ファージ型RNAポリメラーゼ

This research purpose is to define the network of gene expression by communication between organelles and nuclear genomes within a plant cell. The novel findings obtained are as follows.1.We constructed a plastid DNA chip for microarray analysis of plastid genes from the moss, Physcomitrella patens.2.We reported phylogenetic analyses using 51 genes from the entire plastid genome sequences of 20 representative plant species. This indicated that extant bryophytes (mosses, liverworts, and hornworts) form a monophyletic group with high statistical confidence and that extant bryophytes are likely sisters to extant vascular plants. We propose bryophyte monophyly as the current best hypothesis.3.We developed plastid transformation technique for the moss, Physcomitrella patens and constructed stable plastid trnR-CCG knockout moss transformants. The trnR-CCG knockout transformants indicated that the P.patens trnR-CCG gene is not essential for plastid function.4.We identified a nuclear gene, PpR … More poA, encoding the alpha subunit of plastid-encoded plastid RNA polymerase (PEP). We generated and characterized PpRpoA knockout mosses. Primer extension, northern blot, run-on transcription assay and in vitro transcription assays first demonstrated that both PEP and NEP(nuclear-encoded plastid RNA polymerase) exist in the moss and recognize the common promoters of photosynthesis genes and non-photosynthesis genes.5.We identified three PpSig genes encoding plastid sigma factor. Among the three PpSig genes, only PpSig5 was clearly controlled by the circadian clock and/or by blue light signaling in the moss P.patens.6.An extensive survey of the Physcomitrella expressed sequence tag(EST) databases revealed 36 ESTs encoding pentatricopeptide repeat(PPR) motif-containing proteins. We further characterized five full-length cDNAs encoding plastid-localized PPR proteins, PPR513-10 and PPR566-6 were expressed differentially in protonemata grown under different light-dark conditions, suggesting they have distinctive functions in chloroplasts. This is the first report and analysis of genes encoding PPR proteins in bryophytes. Less

这项研究的目的是通过植物细胞内细胞器和核基因组之间的通讯来确定基因表达的网络。所获得的新发现如下：1.我们构建了一个用于绿叶苔藓叶绿体基因芯片的微阵列分析。2.我们报道了来自20个代表性植物物种的51个基因的系统发育分析。这表明现存的苔藓植物(苔藓、地钱和角藻)形成了一个具有很高统计置信度的单系群，并且现存的苔藓植物可能是现存维管植物的姐妹。我们提出了苔藓单子体作为目前最好的假说。3.我们发展了苔藓叶绿体转化技术，构建了稳定的叶绿体trnR-CCG敲除苔藓转化子。TrnR-ccg基因敲除转化子表明，P.patens的trnR-ccg基因对叶绿体的功能不是必需的。4.我们鉴定了一个核基因ppr…更多的POA，编码叶绿体编码的质体RNA聚合酶(PEP)的阿尔法亚单位。我们产生了PpRpoA基因敲除苔藓并对其进行了表征。通过引物延伸、Northern杂交、Run-On转录实验和体外转录实验，首次证明PEP和NEP都存在于苔藓中，识别光合作用基因和非光合作用基因的共同启动子。5.我们鉴定了3个编码体Sigma因子的PpSig基因。在三个PpSig基因中，只有PpSig5在苔藓中明显受生物钟和/或蓝光信号控制。6.对Physcomitrella表达序列标签(EST)数据库的广泛调查发现，36个EST编码含有五肽重复(PPR)基序的蛋白。我们进一步鉴定了5个编码PPR蛋白的全长cDNA，PPR513-10和PPR566-6在不同光暗条件下生长的原生线虫中有不同的表达，表明它们在叶绿体中具有不同的功能。这是首次报道和分析苔藓植物中编码PPR蛋白的基因。较少

项目成果

Nakamura, T., Furuhashi, Y., Hasegawa, K., Hashimoto, H., Obokata, J., Sugita, M., Sugiua, M.: "Array-based analysis on tobacco plastid transcripts : preparation of a genomic microarray containing all genes and intergenic regions"Plant Cell Physiol. 44. 8

Nakamura, T.、Furuhashi, Y.、Hasekawa, K.、Hashimoto, H.、Obokata, J.、Sugita, M.、Sugiua, M.：“基于阵列的烟草质体转录物分析：基因组微阵列的制备

Chloroplast phylogeny indicates that bryophytes are monophyletic.

• DOI: 10.1093/molbev/msh203
• 发表时间: 2004-10
• 期刊: Molecular biology and evolution
• 影响因子: 10.7
• 作者: T. Nishiyama;P. Wolf;M. Kugita;R. Sinclair;M. Sugita;C. Sugiura;T. Wakasugi;Kyoji Yamada;K. Yoshinaga;K. Yamaguchi;K. Ueda;M. Hasebe

Sugiura, C., Kobayashi, Y., Aoki, T., Sugita, C., Sugita, M.: "Complete choloroplast DNA sequence of the moss Physcomitrella patens reveals loss of rpoA from the chloroplast genome : evidence for the loss and relocation of rpoA from the chloroplast to the

Sugiura, C.、Kobayashi, Y.、Aoki, T.、Sugita, C.、Sugita, M.：“苔藓小立碗藓的完整叶绿体 DNA 序列揭示了叶绿体基因组中 rpoA 的丢失：丢失和重新定位的证据

Tanaka, M., Ogawa, N., Ihara, K., Sugiyama, S., Mukohata, Y.: "Cytochrome aa3 and its gene duplication in Haloferax volcanii"J. Bacteriol.. 184. 840-845 (2002)

Tanaka, M.、Okawa, N.、Ihara, K.、Sugiyama, S.、Mukohata, Y.：“Haloferax volcanii 中的细胞色素 aa3 及其基因复制”J.

Yamaguchi, K., Mayfield, S.P., Sugita, M.: "Transcriptional and translational regulation in biosynthesis and repair. In : Photosystem II : The water/plastoquinone oxido-reductase in photosynthesis"Kluwer Publications (in press). (2003)

Yamaguchi, K.、Mayfield, S.P.、Sugita, M.：“生物合成和修复中的转录和翻译调节。见：光系统 II：光合作用中的水/质体醌氧化还原酶”Kluwer Publications（正在出版）。