Amino acid mimicry: Insights into glyphosate transport and toxicity to mitochondria

氨基酸拟态：深入了解草甘膦转运和线粒体毒性

• 批准号: 10573869
• 负责人: Jennifer Gallagher
• 金额: $ 7.6万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-10 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10573869
• 关键词: Acute; Affect; Amino Acids; Animals; Aromatic Amino Acids; Aspartate; Bacteria; Binding; Binding Proteins; Biochemical; Brain; Bypass; Cell Cycle; Cells; Chemical Exposure; Cytoplasm; DNA Damage; Data; Data Set; Diet; Disease; Down-Regulation; Enzymes; Evolution; Food Chain; Food Contamination; Formulation; Future; Genes; Genetic; Glutamate Transporter; Glutamates; Goals; Growth; Health; Herbicides; Human; Human Development; Malate-Aspartate Shuttle Pathway; Malates; Measures; Metabolic Diseases; Metabolism; Mitochondria; Mitochondrial Proteins; Modeling; NADH; Nutrient; Nutrient availability; Occupational Exposure; Organ; Organism; Pathway interactions; Penetration; Plants; Preparation; Regulation; Repression; Reproduction; Research; Resistance; Role; Saccharomyces cerevisiae; Site; Structure; Supplementation; Testing; Time; Tissues; Toxic effect; Transportation; Urine; Yeasts; cancer type; experimental study; exposed human population; fungus; genome wide association study; glyphosate; gut microbiome; innovation; insight; mimicry; mitochondrial metabolism; model organism; mutant; nervous system disorder; permease; sensor; shikimate; surfactant; transcriptomics

Extensive use of glyphosate-based herbicides (GBH) has led to glyphosate entering the food chain, and causing human sera and urine levels to increase over time. This exposure has led to numerous claims that glyphosate causes diseases ranging from multiple types of cancer to affecting human development and reproduction. However, no mechanism of glyphosate import into human cells is known; in yeast, importation occurs through the glutamate/ aspartate (D/E) transporters due to its structural resemblance. Glyphosate is thought not to have acute effects on humans because they lack the shikimate pathway that produces aromatic amino acids (WYF), which is inhibited in the presence of glyphosate, and humans instead acquire aromatic amino acids through diet or the gut microbiome. The differences in commercial preparations of glyphosate have complicated the studies because GBHs have surfactants that increase tissue penetration. Yeast can bypass the inhibition of the shikimate pathway when supplemented with (WYF), which permits the assessment of the role of surfactants or, more likely, discover the unknown glyphosate targets. Our initial studies have found that genes regulating mitochondria, DNA damage, and the cell cycle are differentially regulated in GBH treatments. The long-term goal is to identify the glyphosate transportation mechanisms into cells, the brain, and other tissues that affect mitochondrial metabolism. This application's objective is to determine how mitochondrial metabolism is affected by glyphosate alone and in commercial formulations in the model organism S. cerevisiae. Our central hypothesis is that the transport of glyphosate is due to mimicry of glutamate and aspartate; thus, it will affect other enzymes that utilize glutamate and aspartate, especially within the mitochondria. The rationale of this proposal is that mitochondrial effects of glyphosate have a biochemical basis and will provide a mechanistic understanding of cellular effects in vertebrate species. Specific aims proposed are 1. Measure the import of glyphosate into different compartments in different mutants and how adding D/E rescues growth inhibition from glyphosate 2. Measure changes in specific mitochondrial metabolites (ATP and NADH) in glyphosate treated cells. The proposed research is innovative because the hypothesis proposed is using unbiased experiments, such as genome-wide association, transcriptomics, and In-Lab Evolution experiments to determine the mechanism of extra- and intracellular glyphosate transport using D/E transporters. Glyphosate mimics D/E amino acids in transport, so it likely affects other enzymes that use D/E, particularly in the mitochondria. In plants, fungi, and bacteria, D/E transporters have all been implicated in glyphosate transport and glyphosate affecting mitochondrial functions. While humans do not have the shikimate pathway, they have D/E transporters, and conserved mitochondrial proteins that use D/E, which may be the off-targets of glyphosate leading to the range of diseases claimed to have been caused by glyphosate.

广泛使用基于草甘膦的除草剂（GBH）导致草甘膦​​进入食物链，并且 导致人类血清和尿液水平随时间增加。这种暴露导致了许多主张 草甘膦会导致从多种类型的癌症到影响人类发展和 生殖。然而，没有草甘膦导入到人类细胞中的机制。在酵母中，进口 由于其结构相似，通过谷氨酸/天冬氨酸（D/ E）转运蛋白发生。草甘膦是 认为没有对人类产生急性影响，因为它们缺乏产生芳香的寒冷途径 氨基酸（WYF），在草甘膦存在下被抑制，而人类则获得芳香 通过饮食或肠道微生物组的氨基酸。草甘膦商业制剂的差异 由于GBH的表面活性剂会增加组织穿透性，因此使研究变得复杂。酵母可以 补充（WYF）时，绕过对光滑途径的抑制，这允许评估 表面活性剂的作用或更有可能发现未知的草甘膦靶标。我们的最初研究有 发现调节线粒体，DNA损伤和细胞周期的基因在GBH中受到差异调节 治疗。长期目标是鉴定草甘膦的运输机制，大脑，大脑， 以及其他影响线粒体代谢的组织。该应用程序的目标是确定如何 线粒体代谢受草甘膦的影响，在模型中的商业配方中受到影响 有机体S. cerevisiae。我们的中心假设是草甘膦的运输是由于模仿的 谷氨酸和天冬氨酸；因此，它将影响其他利用谷氨酸和天冬氨酸的酶，尤其是 在线粒体内。该提议的理由是草甘膦的线粒体效应具有 生化基础，将提供对脊椎动物物种细胞作用的机械理解。 提出的具体目的是1。测量草甘膦在不同的隔室中的进口 突变体以及添加D/E如何挽救草甘膦2的生长抑制作用2。衡量特定的变化 草甘膦处理细胞中的线粒体代谢物（ATP和NADH）。拟议的研究是创新的 因为提出的假设是使用公正的实验，例如全基因组关联，所以 转录组学和LAB内进化实验，以确定细胞外和细胞内的机理 使用D/E转运蛋白的草甘膦转运。草甘膦模仿运输中的D/E氨基酸，因此很可能 影响使用D/E的其他酶，尤其是在线粒体中。在植物，真菌和细菌中，D/E 转运蛋白都与草甘膦转运和影响线粒体功能的草甘膦有关。 虽然人类没有阳性途径，但它们具有D/E转运蛋白和保守的线粒体 使用d/e的蛋白质，这可能是草甘膦的非目标，导致声称的疾病范围 是由草甘膦引起的。