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：
8197685
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 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8197685
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 public health relevance 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个重要的机会主义寄生虫，导致在子宫内获得的免疫损害和严重的先天缺陷中威胁生命的感染。寄生虫在专门的室内生长，即感染宿主细胞内的寄生虫液泡（PV）。定义该利基市场的边界是PV膜（PVM），它用作寄生虫和宿主之间的界面。尽管它很重要，但对PVM的调查仍然有限。我们最近完成了对T. gondii PVM的蛋白质组学分析，从而鉴定了123种蛋白质，其中70个蛋白质似乎是新颖的。在这些新型蛋白质中，26被注释为“假设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主题，表明它们确实是新颖的。虽然有几个在毒品上看起来独特，但其他一些人在其他不太易于处理的apicomplexa中也具有类似的注释同源物，包括疟原虫和隐孢子虫属。暗示保守的功能。作为潜在局限于PVM的新型蛋白质，只有在寄生虫感染的背景下才发现细胞器，这些假设的ORF代表了可能用作药物靶标的潜在独特活动。在这项研究中，我们利用了最近开发的技术来表位标签，并将假设的ORF定位为克隆经过验证的基因并尝试敲除基因敲除的前奏。这些研究一起代表了现有数据集作为有限的短期项目的次要分析，其揭示新活动的潜力很高。因此，它非常适合RO3资金机制。通过此提案，我们旨在执行这些孤儿基因的初始表征，以在寄生虫生物学中建立它们的功能。公共卫生相关性：弓形虫弓形虫在免疫受损的个体中导致严重疾病，并且是一种模型的生物体，用于几种实验性较低的寄生虫。这项研究旨在研究真正的新型蛋白质，这些蛋白质被确定为寄生虫液泡膜的组成部分，这是一种独特而必不可少的寄生虫细胞器。我们希望完成这项工作，以开放寄生虫生物学的新调查领域。