Regulation of secondary metabolism and cellular differentiation by A-factor and signal transduction systems in Streptomyces

链霉菌 A 因子和信号转导系统对次生代谢和细胞分化的调节

• 批准号: 04454075
• 负责人: HORINOUCHI Sueharu
• 金额: $ 4.03万
• 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454075/
• 关键词: Streptomyces; A-factor; Antibiotic production; morphogenesis; Signal transduction; Protein phosphorylation

1. A-factor is a microbial hormone controlling streptomycin production and sporulation. An A-factor-dependent promoter, the strR promoter, in the streptomycin biosynthetic gene cluster of S.griseus was identified. Gel retardation assays detected a protein that specifically bound the strR promoter.2. The AfsR protein is essential for the biosynthesis of A-factor, actinorhodin, and undecylprodigiosin in S.coelicolor. The AfsR protein was purified to homogeneity and found to be phosphorylated by a specific kinase. It is conceivable that the extent of phosphorylation of AfsR modulates its regulatory activity which, in turn, regulates expression of some target genes involved in secondary metabolism in this strain.3. In vitro phosphorylation reactions using extracts of S.griseus cells revealed the presence of multiple phosphorylated proteins. Most of these phosphorylations were distinctly inhibited by staurosporine and K-252a which are known to be eucaryotic protein kinase inhibitors. Consistent with this, these kinase inhibitors inhibited antibiotic production and aerial mycelium formation in S.griseus and S.coelicolor.

1. A因子是一种控制链霉素产生和孢子形成的微生物激素。鉴定了灰色链霉菌链霉素生物合成基因簇中的A因子依赖性启动子strR启动子。凝胶阻滞试验检测到特异性结合strR启动子的蛋白质。的AfsR蛋白是必不可少的A-因子，放线菌紫素，和undecylprodigiosin天蓝色链霉菌的生物合成。AfsR蛋白被纯化至均一，并被发现被特定的激酶磷酸化。推测AfsR的磷酸化程度调节其调节活性，进而调节该菌株中参与次级代谢的一些靶基因的表达.在体外磷酸化反应中使用的提取物的S.griseus细胞揭示了存在多个磷酸化的蛋白质。已知的真核蛋白激酶抑制剂staurosporine和K-252 a可明显抑制大多数磷酸化。与此相一致，这些激酶抑制剂抑制抗生素的生产和气生菌丝体的形成在S.griseus和S. coelicolor。

项目成果

S.Horinouchi: "Bioscience and Streptomyces". Igaku-Shuppan Center, Tokyo(in press), (1994)

S.Horinouchi：“生物科学和链霉菌”。

H.Ishizuka: "A putative two-component regulatory system involved in secondary metabolism in Streptomyces spp." Journal of Bacteriology. 174. 7585-7594 (1992)

H.Ishizuka：“一种假定的双组分调节系统，涉及链霉菌属的次生代谢。”

A.Matsumoto: "Phosphorylation by an eukaryotic protein kinase of the AfsR protein involved in secondary metabolism in the bacterial genus Streptomyces" Gene. (in press).

A.Matsumoto：“参与链霉菌属细菌次生代谢的 AfsR 蛋白被真核蛋白激酶磷酸化”基因。

S.Horinouchi: "Regulation of gene expression via protein phosphorylation in Streptomyces" Bioscience & Bioindustry. 50. 425-432 (1992)

S.Horinouchi：“链霉菌中通过蛋白质磷酸化调节基因表达” Bioscience

S.Horinouchi: "Autoregulatory factors and communication in actinomycetes" Annual Review of Microbiology. 46. 377-398 (1992)

S.Horinouchi：“放线菌的自动调节因素和通讯”微生物学年度评论。