Regulation of secondary metabolism and cell differentiation by A-factor and protein phosphorylation in Streptomyces.

链霉菌中 A 因子和蛋白质磷酸化对次生代谢和细胞分化的调节。

• 批准号: 06454073
• 负责人: HORINOUCHI Sueharu
• 金额: $ 4.86万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454073/
• 关键词: Streptomycetes; Utoregulator; Antibiotic; Morphogenesis; Signal transduction; A-ファクター; 抗生物質生産

(1) The A-factor recepor protein (ArpA) was purified from S.griseus and arpA was cloned by PCR.We also cloned two arpA homologs from S.coelicolor A3 (2), which probably controlled antibiotic production in this strain, as determined by the phenotypes of each of disruptants.(2) Transcription of amfR,a regulatory gene for aerial mycelium formation in S.griseus, was controlled by A-factor. A protein able to bind to its promoter region was purified and its gene was cloned.(3) An afsK homolog (afsK2) was cloned from S.griseus. Genetic studies showed that the afsK2 gene is involved in aerial mycelium formation under the conditons of high osmolality(4) Upstream from afsA was present a gene encoding a protein similar to prokaryotic transcriptional factors. The expression of afsA therefore apeared to be controlled by this upstream gene.(5) An A-factor-responsive transcriptional activator (Adp) that bindsto the strR promoter was purified to homogeneity. The molecular mass of Adp was 40 kDa.(6) A tyrosine phosphatase gene was cloned from S.coelicolor A3 (2) as a gene that caused pigmentation in S.lividans. The phosphatase produced in E.coli showed a tyrosine phosphatase activity.

(1)从灰链霉菌中分离纯化了A因子受体蛋白(ARPA)，并用PCR方法克隆了ARPA基因，并从天蓝色链霉菌A3(2)中克隆了两个ARPA同源物，根据各干扰物的表型确定该菌株可能控制了抗生素的产生。纯化了能够与其启动子区域结合的蛋白，并克隆了其基因。(3)从灰色链霉菌中克隆了afsK同源基因(AfsK2)。遗传学研究表明afsK2基因参与了高渗透压条件下气生菌丝的形成。(4)AFSA上游存在一个编码与原核转录因子类似的蛋白质的基因。因此，AFSA的表达似乎受这个上游基因的控制。(5)纯化了与STRR启动子结合的A因子反应转录激活因子(ADP)。(6)从天蓝色链霉菌A3(2)中克隆了一个引起变铅青链霉菌色素沉着的酪氨酸磷酸酶基因。在大肠杆菌中产生的磷酸酶具有酪氨酸磷酸酶活性。

项目成果

A.Matsumoto: "Phosphorylation of the AfsR protein involved in secondary metabolism in Streptomyces spp. by a eukaryotic-type proteinkinase" Gene. 146. 47-56 (1994)

A.Matsumoto：“真核型蛋白激酶对参与链霉菌属次生代谢的 AfsR 蛋白进行磷酸化”基因。

S.Horinouchi: "Dynamic Aspect of Natural Products chemistry-Molecular Biological Approaches" Kodansha-Scientific(印刷中),

S. Horinouchi：“天然产物化学的动态方面 - 分子生物学方法”讲谈社科学（出版中），

Horinouchi, S., and T.Beppu: "Autoregulators. Genetics and Biochemistry of Antibiotic Production. Butterworth-Heinemann Publishers, Boston" 103-119 (1994)

Horinouchi, S. 和 T.Beppu：“自动调节器。抗生素生产的遗传学和生物化学。Butterworth-Heinemann 出版社，波士顿”103-119 (1994)

Kudo, N., M.Kimura, T.Beppu, and S.Horinouchi: "Cloning and characterization of a gene involved in aerial mycelium formation in Streptomyces griseus" J.Bacteriol. 177. 6401-6410 (1995)

Kudo, N.、M.Kimura、T.Beppu 和 S.Horinouchi：“灰色链霉菌气生菌丝体形成相关基因的克隆和表征”J.Bacteriol。

Sueharu Horinouchi: "Signal transduction via A-factor that controls antibiotic production and morphogenesis in Streptomyces." Proceedings of International Symposium on Functional Biomaterials. 88-98 (1995)

Sueharu Horinouchi：“通过 A 因子进行信号转导，控制链霉菌中抗生素的产生和形态发生。”