Nematicidal Chalcones: A Genetic Approach to Uncovering the Action Mechanism

杀线虫查尔酮：揭示作用机制的遗传学方法

• 批准号: 10797915
• 负责人: ALEJANDRO CALDERON-URREA
• 金额: $ 6.5万
• 依托单位: CALIFORNIA STATE UNIVERSITY FRESNO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10797915
• 关键词: Agriculture; Animals; Binding; Caenorhabditis elegans; Cells; Chalcones; DNA Sequence; Enzymes; Genes; Genetic Screening; Health; Human; Infection; Medicine; Meloidogyne; Mutagens; Mutation; Nematoda; Parasites; Parasitic nematode; Plants; Predisposition; Proteins; Resistance; Science; Soil; Testing; Underrepresented Populations; Veterinary Medicine; Work; experience; genetic approach; helminth infection; microorganism; mortality; mutant; novel; recruit; student training; tool; transmission process

Title: Nematicidal chalcones: a genetic approach to uncovering the action mechanism Project Summary/Abstract: Parasitic Nematodes (PNs) are enemy to both plant and animals (including humans). These parasites have a tremendous impact on medicine, veterinary medicine, and agriculture, causing human infections and mortality (see WHO “Soil-transmitted helminth infections” factsheet) and significant annual monetary losses. For example, there is an estimated loss of $157 billion worldwide (Li et al., 2007) due to infestation by plant parasitic nematodes. Therefore, developing new nematicidal agents and understanding their mechanism of action is of paramount importance; it is likely that a solution to nematode control will come from the use of cocktails of nematicidal agents, which can control nematodes in different ways. Previous work in our lab identified two organic chalcones (chalcones 17 and 30) as effective nematicidals on Caenorhabditis elegans and the plant parasitic nematode Meloidogyne incognita (Saeed, et al. 2012, and Calderón-Urrea, et al., unpublished results). Furthermore, these chalcones have no significant effects on either soil microorganisms or human cells (Calderón-Urrea, et al., unpublished results). However, the mechanism of action of these chalcones is not known. It is possible that these chalcones target essential proteins (enzymes), which are rendered infective by the chalcones. If this is the case, alterations in the binding of the chalcones to said proteins can be a powerful tool to elucidate the mechanism of action of the chalcones. We hypothesize that “the susceptibility to chalcones is due to the action of the chalcone on a protein product, perhaps an enzyme, and therefore mutations on the gene (or genes) encoding that protein will render the chalcone action ineffective”. We are testing this hypothesis by conducting a forward genetic screen on mutagenized C. elegans nematodes, and characterizing the isolated mutants. The specific objectives of this project are: 1) To test resistance to chalcones 17 and 30 and identification of mutants overcoming chalcone effects; 2) Characterization of chalcone resistant mutants; and 3) Identification of the DNA sequences involved in the chalcone resistant mutants. The PI will use his extensive experience in recruiting and training students, particularly from underrepresented groups in sciences, to actively engage them in the execution of this project.

标题：杀线虫查耳酮：揭示作用机制的遗传学方法 项目摘要/摘要： 寄生线虫 (PN) 是植物和动物（包括人类）的敌人。 对医学、兽医学和农业产生巨大影响，导致人类感染和死亡 （参见世卫组织“土源性蠕虫感染”情况说明书）以及每年重大的金钱损失。 由于植物寄生虫的侵扰，全球估计损失 1570 亿美元（Li 等人，2007 年） 因此，开发新的杀线虫剂并了解其作用机制至关重要。 最重要的是，控制线虫的解决方案可能来自于使用混合物 我们实验室之前的工作发现了两种杀线虫剂，可以通过不同的方式控制线虫。 有机查尔酮（查尔酮 17 和 30）作为对秀丽隐杆线虫和植物的有效杀线虫剂 寄生线虫根结线虫（Saeed 等人，2012 年；Calderón-Urrea 等人，未发表结果）。 此外，这些查酮对土壤微生物或人体细胞没有显着影响 （Calderón-Urrea 等人，未发表的结果）然而，这些查酮的作用机制尚不清楚。 这些哈酮可能以必需蛋白质（酶）为目标，而这些蛋白质通过 如果是这种情况，改变查耳酮与所述蛋白质的结合可能是一个强大的工具。 阐明查耳酮的作用机制。 由于查耳酮对蛋白质产物（可能是酶）的作用，因此导致基因突变 （或基因）编码该蛋白质将使查尔酮作用无效”。 对诱变的秀丽隐杆线虫进行正向遗传筛选，并表征分离的线虫 该项目的具体目标是： 1) 测试对 halcones 17 和 30 的抗性并进行鉴定。 克服查耳酮效应的突变体；2) 查耳酮抗性突变体的表征；以及 3) 鉴定 PI 将利用他丰富的经验来研究查尔酮抗性突变体所涉及的 DNA 序列。 招募和培训学生，特别是来自科学领域代表性不足的群体的学生，让他们积极参与 在这个项目的执行过程中。