RNAポリメラーゼの金属をとりまく局所の構造と機能

RNA聚合酶中金属周围的局部结构和功能

• 批准号: 09235234
• 负责人: 石浜 明
• 金额: $ 1.34万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09235234/
• 关键词: 転写制御; 転写装置; 蛋白質間相互作用; 蛋白質DNA相互作用; RNAポリメラーゼ; 人工プロテアーゼ; 人工ヌクレアーゼ

大腸菌RNAポリメラーゼは、α_2ββ'σのサブユニット構造をもち、亜鉛を含む金属酵素である。転写調節蛋白因子やDNAの転写調節シグナルとの相互作用による、その機能特異性、特に転写対象遺伝子選択特性変換機構の解明を目的として、各種の転写因子やDNA調節シグナルとの接触実体の解明を目指した。まず分子間接触に異常をもつRNAポリメラーゼ変異体を単離し、変異部位の分析から、RNAポリメラーゼ分子上にそれらの接点を推定した。これら遺伝解析から得られた結果をより直接的に検証する目的で、本年度はふたつのサブユニットα及びσ上の推定接点領域のアミノ酸をシステインに置換し、本研究で開発した核酸及び蛋白質切断活性をもつ化学試薬Fe-BABEを結合した。サブユニット結合Fe-BABEによる接触相手蛋白質またはDNA分子の切断部位を配列分析から決定することに成功した。これらの成果は、転写研究に新たな突破口を開いたのみならず、生体における分子間相互作用解析のために、新たな研究方法を開発した意義としても評価された。

RNA spp., α _ 2 β', β', α _ 2, β, β, α, β, β, The DNA factor is used to determine the interaction characteristics, the mechanical properties, the selected properties, the purpose and the various factors of the protein factor, the DNA factor, the contact factor, the target indicator, the contact factor, the target indicator. Intermolecular contact devices are often used to determine the temperature of the RNA contact, the location analysis, and the presumption of the contact temperature of the RNA contact on the molecule. The results of this study show that the results of this study show that there is no significant difference in the results of this year's study. The results show that the results of the analysis show that the results show that there is no significant difference in the results of the analysis of the results, the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of the analysis of the results show that the results of this year's The alignment analysis of the cut-off site of the DNA molecule was determined by the combination of the Fe-BABE protein and the contact phase of the chiral protein. New breakthroughs in research and research will be launched, the analysis of interactions between molecules in vivo will be conducted, and new research methods will be developed.

项目成果

Greiner,D.P.: "Synthesis of the protein cutting reagent iron (S)-1(p-bromoacetamidebenzym)ethylenediaminetetraacetate and conjucation to cysteine side chains." Bioconjugate Chemistry. 8(1). 44-48 (997)

Greiner，D.P.：“蛋白质切割试剂铁 (S)-1（对溴乙酰胺苯酶）乙二胺四乙酸盐的合成以及与半胱氨酸侧链的缀合。”

Kusano, S.: "Functional interaction of Escherichia coli RNA polymerase with inorganic polyphosphate." Genes to Cells. 2(7). 433-441 (1997)

Kusano, S.：“大肠杆菌 RNA 聚合酶与无机多磷酸盐的功能相互作用。”

Ishihama, A.: "Nucleic Acids & Molecular Biology,Vol.11,Mechanism of Transcription" Springer-Verlag, 327 (1997)

Ishihama, A.：“核酸

Chugani, S.A.: "Activation of the catBCA promoter:probing the interaction of CatR and RNA polymerase through in vitro transcription." J.Bacteriol.179(7). 2221-2227 (1997)

Chugani, S.A.：“catBCA 启动子的激活：通过体外转录探测 CatR 和 RNA 聚合酶的相互作用。”

Ozoline,O.N.: "Specific fluorescent labeling of two functional domains in RNA polymerase alpha subunit." PROTEINS:Structure,Function,and Genetics. 30(1). 1-10 (1997)

Ozoline,O.N.：“RNA 聚合酶 α 亚基中两个功能域的特异性荧光标记。”