{"title":"ActO 是抗生素链霉菌 ZS 中放线菌素生物合成的正向簇定位调节器","authors":"","doi":"10.1016/j.gene.2024.148962","DOIUrl":null,"url":null,"abstract":"<div><div>Actinomycins are a class of cyclic lipopeptide antibiotics produced by <em>Streptomyces</em>, which have rich biological activities and demonstrate great potential value. Among them, actinomycin D is currently the effective drug for some malignant tumor diseases. Although the chemical properties, biological activities and biosynthesis of actinomycins have been extensively studied, the regulation of their biosynthesis remains poorly understood. <em>Streptomyces antibioticus</em> ZS isolated from deep-sea corals is a producer of actinomycin D and actinomycin V. Here, we reported the characterization of a cluster-situated regulator ActO in actinomycins biosynthetic gene cluster (<em>act</em> cluster) of <em>S. antibioticus</em> ZS, which belongs to LmbU family. Deletion of <em>actO</em> completely blocked the synthesis of actinomycins. Overexpression of <em>actO</em> increased the yields of actinomycin D and actinomycin V by 4.4 fold and 2.6 fold, respectively. The result of RT-qPCR showed that ActO activates the transcription of all genes in <em>act</em> cluster. However, no specific binding of His<sub>6</sub>-ActO to the promoters of target genes was observed after electrophoretic mobility shift assay (EMSA). These results proved that ActO serves as a positive regulator involved in the biosynthesis of actinomycins, affecting the transcription of all genes related to the synthesis of intermediates, skeleton modification and extracellular transportation of final products. Moreover, we demonstrated that overexpression of <em>actO</em> is a novel strategy to increase the yields of actinomycins.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6000,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"ActO, a positive cluster-situated regulator for actinomycins biosynthesis in Streptomyces antibioticus ZS\",\"authors\":\"\",\"doi\":\"10.1016/j.gene.2024.148962\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Actinomycins are a class of cyclic lipopeptide antibiotics produced by <em>Streptomyces</em>, which have rich biological activities and demonstrate great potential value. Among them, actinomycin D is currently the effective drug for some malignant tumor diseases. Although the chemical properties, biological activities and biosynthesis of actinomycins have been extensively studied, the regulation of their biosynthesis remains poorly understood. <em>Streptomyces antibioticus</em> ZS isolated from deep-sea corals is a producer of actinomycin D and actinomycin V. Here, we reported the characterization of a cluster-situated regulator ActO in actinomycins biosynthetic gene cluster (<em>act</em> cluster) of <em>S. antibioticus</em> ZS, which belongs to LmbU family. Deletion of <em>actO</em> completely blocked the synthesis of actinomycins. Overexpression of <em>actO</em> increased the yields of actinomycin D and actinomycin V by 4.4 fold and 2.6 fold, respectively. The result of RT-qPCR showed that ActO activates the transcription of all genes in <em>act</em> cluster. However, no specific binding of His<sub>6</sub>-ActO to the promoters of target genes was observed after electrophoretic mobility shift assay (EMSA). These results proved that ActO serves as a positive regulator involved in the biosynthesis of actinomycins, affecting the transcription of all genes related to the synthesis of intermediates, skeleton modification and extracellular transportation of final products. Moreover, we demonstrated that overexpression of <em>actO</em> is a novel strategy to increase the yields of actinomycins.</div></div>\",\"PeriodicalId\":12499,\"journal\":{\"name\":\"Gene\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2024-09-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Gene\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0378111924008436\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"GENETICS & HEREDITY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Gene","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0378111924008436","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
引用次数: 0
摘要
放线菌素是由链霉菌产生的一类环脂肽抗生素,具有丰富的生物活性和巨大的潜在价值。其中,放线菌素 D 是目前治疗某些恶性肿瘤疾病的有效药物。尽管人们对放线菌素的化学性质、生物活性和生物合成进行了广泛的研究,但对其生物合成的调控却知之甚少。从深海珊瑚中分离出的抗生素链霉菌 ZS 是放线菌素 D 和放线菌素 V 的生产者。在此,我们报道了属于 LmbU 家族的抗生素链霉菌 ZS 放线菌素生物合成基因簇(act 簇)中的簇定位调控因子 ActO 的特征。删除 actO 可完全阻止放线菌素的合成。过表达 actO 可使放线菌素 D 和放线菌素 V 的产量分别增加 4.4 倍和 2.6 倍。RT-qPCR 结果显示,ActO 能激活 act 簇中所有基因的转录。然而,在电泳迁移试验(EMSA)中没有观察到 His6-ActO 与目标基因启动子的特异性结合。这些结果证明,ActO 是参与放线菌素生物合成的正调控因子,影响与中间产物合成、骨架修饰和最终产物胞外运输相关的所有基因的转录。此外,我们还证明了过表达 actO 是提高放线菌素产量的一种新策略。
ActO, a positive cluster-situated regulator for actinomycins biosynthesis in Streptomyces antibioticus ZS
Actinomycins are a class of cyclic lipopeptide antibiotics produced by Streptomyces, which have rich biological activities and demonstrate great potential value. Among them, actinomycin D is currently the effective drug for some malignant tumor diseases. Although the chemical properties, biological activities and biosynthesis of actinomycins have been extensively studied, the regulation of their biosynthesis remains poorly understood. Streptomyces antibioticus ZS isolated from deep-sea corals is a producer of actinomycin D and actinomycin V. Here, we reported the characterization of a cluster-situated regulator ActO in actinomycins biosynthetic gene cluster (act cluster) of S. antibioticus ZS, which belongs to LmbU family. Deletion of actO completely blocked the synthesis of actinomycins. Overexpression of actO increased the yields of actinomycin D and actinomycin V by 4.4 fold and 2.6 fold, respectively. The result of RT-qPCR showed that ActO activates the transcription of all genes in act cluster. However, no specific binding of His6-ActO to the promoters of target genes was observed after electrophoretic mobility shift assay (EMSA). These results proved that ActO serves as a positive regulator involved in the biosynthesis of actinomycins, affecting the transcription of all genes related to the synthesis of intermediates, skeleton modification and extracellular transportation of final products. Moreover, we demonstrated that overexpression of actO is a novel strategy to increase the yields of actinomycins.
期刊介绍:
Gene publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses.