Function and Trafficking of Flagellar Membrane Proteins in Leishmania mexicana

墨西哥利什曼原虫鞭毛膜蛋白的功能和运输

基本信息

批准号：
10632896
负责人：
Scott M Landfear
金额：
$ 68万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-22 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10632896
关键词：
Address Affinity Chromatography Back Biochemical Biological Biological Process Biology Cells Collection Cytokinesis Cytoskeletal Proteins Data Dedications Destinations Disease Environment Flagella Glucose Transporter Immune response Immunity In Vitro Individual Infection Insect Vectors Insecta Integral Membrane Protein Knowledge Leishmania Leishmania mexicana Life Cycle Stages Mass Spectrum Analysis Mediating Membrane Membrane Protein Traffic Membrane Proteins Modeling Molecular Mus Organelles Parasites Parasitology Partner in relationship Pathway interactions Persons Phenotype Phlebotominae Play Proteins Protozoa Research Role Sand Flies Sorting Surface Testing Time Trypanosoma brucei brucei Trypanosoma cruzi Vacuole Virulence cell motility cohort disorder control extracellular extracellular vesicles frontier hemoflagellate in vivo infancy knockout gene mutant novel parasitism particle trafficking transmission process vector

项目摘要

Leishmania and other kinetoplastid parasites cause devastating diseases that afflict millions of people. All kinetoplastid parasites contain a whip-like organelle, the flagellum, in all life cycle stages. Studies over decades have underscored the vital and diverse roles that flagella play in the biology of these parasites, including motility, attachment to the gut of insect vectors, sensing the environment, mediating mating, interacting with internal membranes of mammalian host cells, evading the host immune response, etc. As such, flagella are essential for both parasite colonization of and transmission through the insect vector and for survival and virulence inside the vertebrate host. While internal components of the flagellum, such as the axoneme, have been studied in greater detail, the surface of the flagellar membrane (FM) is poorly understood, despite the fact that the FM mediates most of the above biological functions. To better understand the FM and its role in parasitism, this proposal will investigate: Aim 1) how specific FM proteins traffic selectively to this discrete component of the surface membrane; Aim 2) what the biological functions are for several selected FM proteins. The project is founded on initial studies on the KHARON protein, that mediates FM trafficking of a flagellar glucose transporter GT1, a trafficking machine that is essential for viability of intracellular disease- causing amastigote stages. A molecular mechanism for KHARON in delivering GT1 to Intraflagellar Transport particles for trafficking into the flagellum will be tested. Since most FM proteins do not depend upon KHARON for FM trafficking, other FM trafficking machines that interact with those FM proteins will be identified by tandem affinity purification/mass spectrometry. These novel FM trafficking machines will then be investigated, by gene knockout approaches, for critical functions in both the insect vector and the mammalian infective stages of the life cycle. Three selected FM proteins with known or likely biochemical functions will then be investigated, by gene knockout, for functions in the insect vector and mammalian host stages. These studies will reveal how specific molecular components of the FM mediate critical functions for the parasite in both the insect vector and the mammalian host.

利什曼尼亚和其他动力质体寄生虫会引起毁灭性疾病，折磨数百万人。全部在所有生命周期阶段，动型寄生虫都包含一个类似鞭子的细胞器，鞭毛。研究数十年来强调了鞭毛在这些寄生虫的生物学中所扮演的重要和多种角色，包括运动性，依恋昆虫向量的肠道，传感环境，介导交配，与哺乳动物宿主细胞的内部膜相互作用，逃避宿主免疫反应等。这样的鞭毛对于通过昆虫载体的寄生虫定植和传播以及脊椎动物宿主内的生存和毒力。而鞭毛的内部组件，例如轴突已更详细地研究了，鞭毛膜（FM）的表面知之甚少，尽管FM介导了上述大多数生物学功能。更好地了解FM和该提议在寄生虫中的作用将调查：目标1）特定的FM蛋白如何有选择地交通表面膜的离散成分；目标2）几种选定的FM的生物学功能是什么蛋白质。该项目建立在有关Kharon蛋白的初步研究基础上，该研究介导了A 鞭毛葡萄糖转运蛋白GT1，这是一种对细胞内疾病的生存能力至关重要的运输机器 - 引起amastigote阶段。 Kharon在将GT1传递到flagellar转运的分子机制将测试用于运输到鞭毛的颗粒。由于大多数FM蛋白不依赖Kharon 对于FM贩运，其他与这些FM蛋白质相互作用的FM运输机将由串联亲和力纯化/质谱法。这些新颖的FM贩运机器将被调查，通过基因敲除方法，用于昆虫载体和哺乳动物感染力的关键功能生命周期的阶段。然后将三个选定的具有已知或可能生化功能的FM蛋白通过基因敲除研究了昆虫载体和哺乳动物宿主阶段的功能。这些研究将揭示FM的特定分子成分如何介导寄生虫的关键功能昆虫载体和哺乳动物宿主。