Artemisinin Biosynthesis: Control of Transcription

青蒿素生物合成：转录控制

• 批准号: 7934255
• 负责人: PAMELA J WEATHERS
• 金额: $ 6.94万
• 依托单位: WORCESTER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7934255
• 关键词: Abbreviations; Acids; Affect; Anabolism; Antimalarials; Artemisia annua; Artemisinins; Carbon; Complex; Cytochrome P450; D-xylulose-5-phosphate; Data; Development; Dose; Engineering; Enzymes; FPS-FES Oncogene; Flowers; Fructose; Gene Expression; Genes; Genetic Transcription; Geranyltranstransferase; Glucose; Goals; Growth; Hydroxymethylglutaryl-CoA reductase; Individual; Knowledge; Lactones; Malaria; Metabolic; Molecular; Pathway interactions; Pharmaceutical Preparations; Plants; Production; Protein Isoforms; RNA Interference; Regulation; Regulator Genes; Relative (related person); Role; Seedling; Sesquiterpenes; Source; Squalene Synthetase; Staging; Sterols; Sucrose; Terpenes; Terpenoid Biosynthesis Pathway; Testing; Therapeutic; Transcriptional Regulation; Transgenic Organisms; Vascular Plant; Weight; Work; amorpha-4,11-diene synthase; anti-cancer therapeutic; antimicrobial; arteannuin B; artemisic acid; artemisinine; cost; drug production; farnesyl pyrophosphate; gibberellic acid; isopentenyl pyrophosphate; mevalonate; novel; sugar; tool; trichodiene synthase

DESCRIPTION (provided by applicant): Artemisinin (AN), and its derivatives are proven effective antimalarial therapeutics that also been shown to have effective activity as broad spectrum antimicrobials, and anticancer therapeutics. AN is a complex compound and still best produced by the plant Artemisia annua. Little is known, however, about how AN biosynthesis is regulated, although the following is known about its production: more AN or its precursor, artemisinic acid (AA), is made when plants shift from vegetative growth to flowering, or grown on glucose instead of sucrose (our recent data). Since these conditions strongly affect AN production, we propose to use them to study the transcriptional regulation of AN biosynthetic genes. OUR HYPOTHESES: 1. Artemisinin biosynthesis is regulated at multiple steps in the pathway, both early (pre- FDP) and late (post-FDP). 2. Developmental stage and carbon source regulate the transcription of several genes in this pathway. 3. Coordinate control of artemisinin and sterol production occurs at the level of transcription. TECHNICAL OBJECTIVES: The long-term goal of our work is to develop a fundamental understanding of the regulatory control of terpene production in Artemisia annua so that metabolic manipulation of terpenoid biosynthesis can eventually be developed not only for artemisinin but other therapeutically important plant produced terpenoid drugs. This work has 3 objectives to be accomplished over 2.5 years: 1. To compare the relative expression of the terpenoid biosynthetic genes pre farnesyl diphosphate (FDP), and post FDP leading to AN with levels of both AA, DHAA, and AN production in A. annua seedlings as they are shifted from vegetative growth to conditions that induce flowering. 2. To compare the relative expression of the terpenoid biosynthetic genes pre farnesyl diphosphate (FDP), and post FDP leading to AN with levels of both AA, DHAA, and AN production in A. annua seedlings grown in glucose, fructose, and sucrose. 3. Include analysis of SQS expression in these studies to determine coordinate control of expression. Unfortunately little is still known about how important plant products like AN are produced in the plant and high production of terpenoids in low cost field-grown plants still remains challenging. This information is important because there has been little comprehensive effort to fully understand the regulation of not only this very important therapeutic sesquiterpene, but also other plant-derived terpenes that could have potentially very beneficial medicinal applications. The most potent therapeutic available for curing malaria, artemisinin, is produced in the plant, "Sweet Annie", but at levels too low to be able to provide adequate doses to annually treat the millions of cases that occur worldwide. The proposed study is a first step in helping us understand how to control artemisinin's biosynthesis to ultimately increase the production of this drug, and others that are chemically similar, in low cost field-grown plants.

描述（由申请人提供）：青蒿素（AN）及其衍生物被证明是有效的抗疟疗法，也被证明具有作为广谱抗菌剂和抗癌疗法的有效活性。 AN 是一种复杂的化合物，最好由青蒿植物产生。然而，关于 AN 的生物合成是如何调节的，人们知之甚少，尽管对其产生的了解如下：当植物从营养生长转向开花时，或在葡萄糖生长时，会产生更多的 AN 或其前体青蒿酸 (AA)。蔗糖（我们最近的数据）。由于这些条件强烈影响 AN 的产生，我们建议利用它们来研究 AN 生物合成基因的转录调控。我们的假设： 1. 青蒿素生物合成在该途径的多个步骤中受到调节，包括早期（FDP 前）和晚期（FDP 后）。 2.发育阶段和碳源调节该途径中多个基因的转录。 3. 青蒿素和甾醇产生的协调控制发生在转录水平。技术目标：我们工作的长期目标是对青蒿中萜烯生产的调节控制有一个基本的了解，以便最终不仅可以为青蒿素而且可以为其他治疗上重要的植物产生的萜类药物开发萜类生物合成的代谢操纵。这项工作有 3 个目标需要在 2.5 年内完成： 1. 比较法呢基二磷酸 (FDP) 前和 FDP 后导致 AN 的萜类生物合成基因的相对表达与 A 中 AA、DHAA 和 AN 的产生水平青蒿幼苗从营养生长转变为诱导开花的条件。 2.比较在葡萄糖、果糖和蔗糖中生长的青蒿幼苗中，法呢基二磷酸(FDP)前和FDP后导致AN的萜类生物合成基因的相对表达与AA、DHAA和AN产量的水平。 3. 在这些研究中包括 SQS 表达分析，以确定表达的协调控制。不幸的是，人们对于植物中生产AN等植物产品的重要性仍然知之甚少，并且在低成本的田间种植植物中高产萜类化合物仍然具有挑战性。这些信息很重要，因为几乎没有全面的努力来充分了解这种非常重要的治疗性倍半萜烯的调节，而且还没有充分了解可能具有潜在非常有益的医学应用的其他植物源萜烯的调节。治疗疟疾最有效的药物青蒿素是在“甜蜜安妮”植物中产生的，但其含量太低，无法提供足够的剂量来治疗每年全世界发生的数百万病例。这项拟议的研究是帮助我们了解如何控制青蒿素生物合成以最终提高这种药物以及其他化学相似药物在低成本田间种植植物中的产量的第一步。