Relationship of Antibiotic Production and Development in Streptomyces coelicolor

天蓝色链霉菌抗生素生产与发育的关系

• 批准号: 7939390
• 负责人: Amy Gehring
• 金额: $ 33.85万
• 依托单位: WILLIAMS COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939390
• 关键词: Affect; Antibiotics; Antifungal Agents; Antineoplastic Agents; Bacteria; Biological Factors; Calcium; Cells; Complex; Cues; Data; Defect; Development; Developmental Gene; Engineering; Enzymes; Event; Generations; Genes; Goals; Growth; Immunosuppressive Agents; Infection; Knowledge; Life; Life Cycle Stages; Link; Malignant Neoplasms; Medical; Membrane Proteins; Metabolic; Metabolism; Microarray Analysis; Modeling; Molecular; Mutation; Mycelium; Organism; Pentosephosphate Pathway; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacologic Substance; Physiological; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; Regulation; Regulon; Reproduction spores; Research; Sigma Factor; Soil; Source; Streptomyces; Streptomyces coelicolor; Surface; Testing; Time; Transferase Gene; base; biological adaptation to stress; enzyme activity; extracellular; fighting; mutant; overexpression; phosphopantetheinyl transferase; programs; public health relevance

DESCRIPTION (provided by applicant): The broad objective of this proposal is to elucidate some of the molecular connections between the complex, interrelated processes of antibiotic production and morphological development in the model bacterium Streptomyces coelicolor. The streptomycetes are an incredibly valuable source of bioactive molecules, including the majority of antibiotics clinically used. These mycelial bacteria typically synthesize antibiotics and other secondary metabolites concurrently with broader developmental events that result in the generation of an aerial mycelium on the colony surface whose filamentous cells ultimately sporulate. The aims of this study concern two particular types of proteins that influence both antibiotic production and development in S. coelicolor. First, the phosphopantetheinyl transferase are enzymes that catalyze a posttranslational modification required for the activity of many antibiotic biosynthetic enzymes. Mutation of the SCO6673 PPTase gene in S. coelicolor results in loss of the ability to produce calcium dependent-antibiotic but also conversely hyperproduction of the potential anti-cancer molecule undecylprodigiosin. Sporulation is also delayed in this mutant. In order to understand the basis for these diverse phenotypic effects, real-time PCR will be used to assess any differences in expression of known antibiotic regulatory and developmental genes in the PPTase mutant as compared to the wild type. This data will inform a subsequent targeted mutational strategy to engineer S. coelicolor for maximum undecylprodigiosin production. The second aim focuses on understanding how activity of a likely stress response ECF sigma factor, ?U, can block both polyketide antibiotic production and aerial mycelium formation in S. coelicolor. A combination of microarray analysis and proteomic studies comparing wild type S. coelicolor to a rsuA mutant with high, unregulated ?U activity will establish the complete regulon of ?U-dependent genes as well as those indirectly affected by ?U activity. The relevance of the observed elevation of both extracellular protease and pentose phosphate pathway enzyme activity in the rsuA mutant to its antibiotic production and developmental defects will be tested by mutation or overexpression of the relevant genes and analysis of the resulting strains. Mutational analysis will also be used to test the involvement of the likely membrane protein SCO4110 in the regulation of ?U activity. A better understanding of how secondary metabolism and development are mutually influenced may ultimately suggest generalizable strategies for effective pharmaceutical production in the streptomycetes. PUBLIC HEALTH RELEVANCE: The goal of the proposed research is to understand how Streptomyces bacteria make useful compounds, such as antibiotics, as part of their overall life cycle. The better we understand how these beneficial bacteria grow, the more likely it is that we can engineer that growth to produce life-saving drugs for fighting infections and cancers.

描述（由申请人提供）：本提案的主要目的是阐明模型细菌天蓝色链霉菌中抗生素生产和形态发育的复杂、相互关联的过程之间的一些分子联系。链霉菌是生物活性分子的一个非常有价值的来源，包括临床上使用的大多数抗生素。这些菌丝体细菌通常合成抗生素和其它次级代谢物，同时伴随更广泛的发育事件，导致在菌落殖民地表面上产生气生菌丝体，其丝状细胞最终形成孢子。这项研究的目的是关注两种特殊类型的蛋白质，影响抗生素的生产和发展，在S。天蓝色。首先，磷酸泛酰巯基乙胺基转移酶是催化许多抗生素生物合成酶活性所需的翻译后修饰的酶。S.腔肠菌导致产生钙依赖性抗生素的能力丧失，而且相反地导致潜在抗癌分子十一烷基灵菌红素的过度产生。在这种突变体中孢子形成也延迟。为了理解这些不同表型效应的基础，将使用实时PCR评估与野生型相比，PPTase突变体中已知抗生素调控和发育基因表达的任何差异。这些数据将为后续的靶向突变策略提供信息，以工程化S。最大的十一烷基灵菌红素生产。第二个目标的重点是了解如何活动的一个可能的压力反应ECF西格玛因素，？U能抑制聚酮抗生素的产生和气生菌丝体的形成。天蓝色。结合微阵列分析和蛋白质组学研究比较野生型S。coelicolor到rsuA突变体高，不受管制？U活动将建立完整的调节子？U依赖基因以及那些间接受？U活动。将通过相关基因的突变或过表达和所得菌株的分析来测试在rsuA突变体中观察到的胞外蛋白酶和戊糖磷酸途径酶活性升高与其抗生素产生和发育缺陷的相关性。突变分析也将被用来测试参与可能的膜蛋白SCO4110的调节？U活动。更好地了解次生代谢和发展是如何相互影响的，最终可能会建议在链霉菌有效的药物生产的推广策略。 公共卫生关系：拟议研究的目标是了解链霉菌细菌如何制造有用的化合物，如抗生素，作为其整个生命周期的一部分。我们越了解这些有益细菌的生长方式，我们就越有可能设计这种生长来生产对抗感染和癌症的救命药物。