Novel activities and the T. gondii vacuolar membrane

新活性和弓形虫液泡膜

基本信息

批准号：
8029051
负责人：
ANTHONY P. SINAI
金额：
$ 7.43万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-12-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8029051
关键词：
Acute Address Affinity Animal Model Antigens Apicomplexa Area Biochemical Bioinformatics Biological Biological Process Biology Cells Cellular biology Cloning Complex Congenital Abnormality Coupled Cryptosporidium Cyst Cytoskeleton Data Set Development Drug Delivery Systems Employment Endoplasmic Reticulum Epitopes Expression Library Fibroblasts Funding Mechanisms Gene Targeting Genes Genome Grant Heart Homologous Gene Immune Immune Sera In Situ Individual Infection Investigation Length Life Manuscripts Membrane Methods Mitochondria Molecular Molecular Biology Nutrient Open Reading Frames Organelles Orphan Oryctolagus cuniculus Parasites Plasmodium Preparation Proteins Proteome Proteomics Reagent Resources Rewards Risk Role Screening procedure Signal Transduction System Technology Toxoplasma Toxoplasma gondii Vacuole Work base biological systems cDNA Expression gene function genome-wide in utero insight interest knockout gene novel parasite genome pathogen stem

项目摘要

DESCRIPTION (provided by applicant): The protozoan parasite Toxoplasma gondii is an important opportunistic parasite causing life threatening infections in the immune compromised and severe birth defects when acquired in utero. The parasite grows within a specialized compartment, the parasitophorous vacuole (PV) within the infected host cell. The boundary defining this niche is the PV membrane (PVM) that serves as the interface between the parasite and host. Despite its importance, investigation of the PVM has been limited. We have recently completed a proteomic analysis of the T. gondii PVM resulting in the identification of 123 proteins of which 70 appear to be novel. Of these novel proteins, 26 are annotated as "hypothetical ORF's" precluding any assignment of function. As hypothetical ORF's, these loci lack any identifiable motif indicating they are truly novel. While several appear unique to Toxoplasma, others have similarly annotated homologs in other less tractable Apicomplexa including Plasmodium and Cryptosporidium spp. suggesting conserved function. As novel proteins that are potentially localized to the PVM, an organelle found only in the context of parasite infection, these hypothetical ORF's represent potentially unique activities that may serve as drug targets. In this study we exploit recently developed technologies to epitope tag and localize the hypothetical ORF's as a prelude to cloning the validated genes and attempting the gene knockouts. Together these studies represent a secondary analysis of an existing data set as a limited short term project with a high potential of revealing novel activities. As such it is ideally suited to the RO3 funding mechanism. With this proposal we aim to perform the initial characterization of these orphan genes toward establishing their functions in parasite biology. PUBLIC HEALTH RELEVANCE: Toxoplasma gondii causes serious illness in immune compromised individuals and is a model organism for several less experimentally tractable parasites. This study aims to investigate truly novel proteins that were identified as being components of the parasitophorous vacuole membrane, a unique and essential parasite organelle. We expect with the completion of this work to open new areas of investigation in parasite biology.

描述（由申请人提供）：原生动物寄生虫弓形虫是一种重要的机会性寄生虫，当在子宫内获得时，会导致免疫受损和严重出生缺陷的危及生命的感染。寄生虫在一个特殊的隔室中生长，即受感染宿主细胞内的寄生液泡（PV）。定义这个生态位的边界是光伏膜（PVM），它充当寄生虫和宿主之间的界面。尽管 PVM 很重要，但对其的研究仍然有限。我们最近完成了弓形虫 PVM 的蛋白质组学分析，鉴定出 123 种蛋白质，其中 70 种似乎是新的。在这些新蛋白质中，有 26 个被注释为“假设的 ORF”，排除了任何功能分配。作为假设的 ORF，这些基因座缺乏任何可识别的基序，表明它们确实是新颖的。虽然其中一些似乎是弓形虫所独有的，但其他一些在其他不易处理的顶复门（包括疟原虫和隐孢子虫属）中也有类似注释的同源物。表明功能保守。作为可能定位于 PVM（一种仅在寄生虫感染情况下发现的细胞器）的新型蛋白质，这些假设的 ORF 代表了可作为药物靶点的潜在独特活性。在这项研究中，我们利用最近开发的技术来标记表位并定位假设的 ORF，作为克隆验证基因并尝试基因敲除的前奏。这些研究共同代表了对现有数据集的二次分析，作为一个有限的短期项目，具有揭示新活动的巨大潜力。因此，它非常适合 RO3 融资机制。通过这项提议，我们的目标是对这些孤儿基因进行初步表征，以确定它们在寄生虫生物学中的功能。公共卫生相关性：弓形虫会导致免疫功能低下的个体患严重疾病，并且是几种实验上不易处理的寄生虫的模式生物。这项研究旨在研究真正新颖的蛋白质，这些蛋白质被确定为寄生液泡膜的组成部分，寄生液泡膜是一种独特且重要的寄生虫细胞器。我们期望随着这项工作的完成，开辟寄生虫生物学研究的新领域。