Joint Research on Differentiation and Growth specificity of Plant Cells

植物细胞分化和生长特异性联合研究

• 批准号: 11694196
• 负责人: TAKAHASHI Hideo
• 金额: $ 6.34万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694196/
• 关键词: Arabidopsis thallana; chloroplast; sigma factor; telomerase; TERT gene; double strand breaks; nuclear gene; イネ; Ku70-Ku80; 高等植物; AtTERT遺伝子; テロメラーゼ活性; 核コードプラスチドの因子; テロメラーゼ活性阻害; DNAのメチル化; 植物細胞; タバコ培養細胞; テロメラーゼ; S期特異的活性化; テロメラーゼ阻害因子; At T

(1)S phase-specific and auxin-enhanced activation or telomerase was demonstrated using cell-extract of cynchronized tobacco culture cells. Genes (cDNAs) for the catalytic subunits of Arabidopsis thallana and Oryza sativa (AtFERT and OsTERT) were cloned for the first time from higher plants. Highest ArTERT expression in the apical meristems was verified using northern and in situ hybridization. (2) A proteinaceous fraction having an inhibitory activity against telomerase was found in plant cell extracts. A 40 kDa protein presumably involved in the regulation of telomerase activity was identified. (3) Genes (cDNAs) for Ku70 and Ku80 homologs of Arabidopsis thaliana (AtKu70 and AtKu80) were cloned and sequenced. A heterodimer specifically formed between purified AtKu70 and AtKu80 proteins revealed sDNA-dependent ATPase and ATP-dependent DNA helicase as well as specific binding to double-stranded DNA ends. DSBs (double-strand breaks) by bleomycin or MMS treatment of cultured cells induce the expression of these genes. (4) Six genes for the nuclear-encoded plastid sigma factors of Arabidopsis thaliana (originally named SIGA〜SIGF and later renamed SIGI〜SIG6, respectively) were identified. A T-DNA inserted mutant (sig 2-1) in SIG2 (SIGB) gene having pale-green phenotype was isolated. Several plastid-encoded tRNAs including trnE-UUC indispensable for chiorophyII (tetrapyrrole) biosynthesis in plants were drastically reduced but that some tRNAs were not affected so much by the SIG2-deficiency.

(1)利用同步化烟草培养细胞的细胞提取液，证实了S对端粒酶的激活和生长素的促进作用。首次从高等植物中克隆了拟南芥和水稻催化亚基基因(AtFERT和OsTERT)。Northern和原位杂交证实ArTERT在顶端分生组织中表达最高。(2)在植物细胞提取物中发现了一种对端粒酶具有抑制活性的蛋白质组分。鉴定出一个40 kDa的蛋白，可能与端粒酶活性的调节有关。(3)克隆了拟南芥(AtKu70和AtKu80)Ku70和Ku80同源基因(CDNAs)。在纯化的AtKu70和AtKu80蛋白之间形成的异源二聚体显示了sDNA依赖的ATPase和ATP依赖的DNA解旋酶以及与双链DNA末端的特异性结合。博莱霉素或MMS处理培养细胞的双链断裂诱导这些基因的表达。(4)鉴定了拟南芥核编码的6个胞质sigma因子基因(最初命名为sigA~sigf，后更名为sig1~sig6)。分离到一个淡绿色的SIG2基因T-DNA插入突变体(sig2-1)。包括植物叶绿体II(四吡咯)生物合成所必需的trnE-UUC在内的几个叶绿体编码的tRNAs显著减少，但一些tRNAs没有受到SIG2缺乏的太大影响。

项目成果

Kanamaru, K. et al.: "Plastidic RNA polymerase a factors in Arabidopsis"Plant Cell Phvsiol.. 40. 832-842 (1999)

Kanamaru，K.等人：“拟南芥中质体RNA聚合酶a因子”Plant Cell Phvsiol.. 40. 832-842 (1999)

J.Grelihuber et al.: "Origin of facultative heterochromatin in the endosperm of Gagea lutea (Liliaceae)"Protoplasma. 212. 217-226 (2000)

J.Grelihuber 等人：“Gagea utea（百合科）胚乳中兼性异染色质的起源”原生质体。

Kanamaru, K. et al: "Chloroplast Targeting, Distribution and transcriptional fluctuation of AtMinD1, a eubacterial-type factor criticdal for chloroplast division"Plant Cell Physiol. 41(10). 1119-1128 (2000)

Kanamaru, K. 等人：“AtMinD1 的叶绿体靶向、分布和转录波动，这是叶绿体分裂的真细菌型关键因子”Plant Cell Physiol。

Kejnovsky e. et al: "Localization of male-specifically expressed MROS genes of Silene latifolia by PCR on flow-sorted sec chrfomosomes and autosomes"Genetics. 158. 1269-1277 (2001)

科伊诺夫斯基E.

Kejnovsky E. et al.: "Localization of male-specifically expressed MROS genes of Silene latifolia by PCR on flow-sorted sex chromosome and autosomes"Genetics. 158. 1269-1277 (2001)

Kejnovsky E. 等人：“通过流式分选性染色体和常染色体上的 PCR 定位 Silene latifolia 雄性特异性表达的 MROS 基因”遗传学。