Nematicidal Chalcones: A Genetic Approach to Uncovering the Action Mechanism

杀线虫查耳酮：揭示作用机制的遗传学方法

• 批准号: 10608187
• 负责人: ALEJANDRO CALDERON-URREA
• 金额: $ 13.4万
• 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/10608187
• 关键词: Agriculture; Animals; Binding; Caenorhabditis elegans; Cell Death; Cells; Chalcones; DNA Sequence; Enzymes; Genes; Genetic Complementation Test; Genetic Screening; Goals; Health; Human; Infection; Maps; Medicine; Meloidogyne; Molecular; Mutagens; Mutation; Nematoda; Parasites; Parasitic nematode; Pathway interactions; Phenotype; Plants; Predisposition; Principal Investigator; Proteins; Resistance; Science; Soil; Testing; Underrepresented Populations; Veterinary Medicine; Work; cross immunity; deep sequencing; experience; genetic approach; helminth infection; microorganism; mortality; mutant; novel; programs; recruit; response; screening; student training; tool; transmission process; uptake

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elegans and the plant parasitic nematode Meloidogyne incognita (Saeed, et al. 2012, and Calderón-Urrea, et al., unpublished results).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these chalcones have no significant effects on either soil microorganisms or human cells (Calderón-Urrea, et al., unpublished results).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：杀线虫查尔酮：揭示作用机制的遗传方法 项目概要/摘要： 寄生线虫是植物和动物（包括人类）的天敌。这些寄生虫 对医学、兽医学和农业产生巨大影响，导致人类感染和死亡 (see世卫组织“土壤传播的蠕虫感染”概况介绍）和每年重大的金钱损失。比如说， 全世界估计损失1570亿美元（Li等人，2007年，由于植物寄生虫的感染， 线虫因此，开发新的杀线虫剂并了解其作用机制是至关重要的。 最重要的是，线虫控制的解决方案很可能来自于使用以下混合物： 杀线虫剂，其可以不同方式控制线虫。我们实验室以前的工作确定了两个 有机查耳酮（查耳酮17和30）作为对秀丽隐杆线虫和植物有效的杀线虫剂 寄生线虫南方根结线虫（Saeed等人，2012年，和Calderón-Urrea等人，未公布的结果）。 此外，这些查耳酮对土壤微生物或人体细胞没有显著影响 （Calderón-Urrea等人，未公布的结果）。然而，这些查尔酮的作用机制尚不清楚。 这些查耳酮可能靶向必需蛋白质（酶），这些必需蛋白质（酶）通过蛋白质（酶）而被赋予感染性。 查耳酮如果是这种情况，改变查耳酮与所述蛋白质的结合可能是一个强有力的工具 阐明查耳酮的作用机制。我们假设“对查耳酮的易感性是 由于查耳酮对蛋白质产物的作用，可能是一种酶，因此基因突变 (or编码该蛋白质的基因）将使查耳酮作用无效”。我们正在测试这个假设， 对诱变的C. elegans线虫，并表征分离的 变种人本项目的具体目标是：1）检测对查耳酮17和查耳酮30的抗性， 突变体克服查耳酮效应; 2）查耳酮抗性突变体的表征;和3）鉴定 查耳酮抗性突变体的DNA序列PI将利用其丰富的经验， 招募和培训学生，特别是来自科学领域代表性不足群体的学生，使他们积极参与 在这个项目的执行中。