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Transcriptional Regulators of the Malarial var Multigene Family

疟疾变种多基因家族的转录调节因子

基本信息

批准号：
8088124
负责人：
CHOUKRI BEN MAMOUN
金额：
$ 20.7万
依托单位：
YALE UNIVERSITY
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：
2010-06-15 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8088124
关键词：
Affinity Chromatography Antibodies Antigenic Variation Binding Biochemical Biological Cell Nucleus Cells Cellular biology Cessation of life Chromosomes Clinical Collaborations DNA Development Drug usage Epitopes Erythrocytes Experimental Designs Fluorescent in Situ Hybridization Gene Expression Genes Genetic Techniques Genome Goals Human Immune response Immunochemistry In Vitro Infection Invaded Laboratories Malaria Mass Spectrum Analysis Membrane Proteins Methodology Methods Molecular Molecular Biology Multigene Family Names Nucleic Acid Regulatory Sequences Nutrient Parasites Plasmodium falciparum Plasmodium falciparum erythrocyte membrane protein 1 Play Population Principal Investigator Promoter Regions Protein Binding Protein C Proteins Proteome Reagent Regulation Repression Research Research Personnel Research Project Grants Resistance Role Site Staining method Stains TEV protease Tetanus Helper Peptide Time Transcription Coactivator Transcription Repressor/Corepressor Transfection Transgenic Organisms Variant Virulence Virulence Factors chromatin immunoprecipitation crosslink in vivo interest link protein novel strategies promoter public health relevance tool transcription factor uptake vector

项目摘要

DESCRIPTION (provided by applicant): The goal of this research project is to develop a new approach that combines chromatin immunoprecipitation, tandem affinity purification and mass spectrometry to identify possible activators and/or repressors of the var multigene family encoding the malaria virulence factor, Plasmodium falciparum erythrocyte membrane protein 1, PfEMP1. During their intraerythrocytic development, malaria parasites switch expression to alternative forms of this variant surface protein to evade the host immune response. The genome of each P. falciparum parasite contains approximately 60 var genes distributed on different chromosomes. However, only one var gene is expressed by each parasite in a mutually exclusive manner. The promoter region of var genes plays an important role in this regulation and it is believed that this mechanism might involve specialized repressor (PfVPR) and/or activator (PfVPA) molecules. The specific aims of the current proposal include experimental designs and methods, some newly developed in our laboratories for P. falciparum such as tandem affinity purification and fluorescent in situ hybridization (FISH). They are: Aim I. To identify PfVPA and PfVPR regulators of var gene expression by chromatin immunoprecipitation, tandem affinity purification and mass spectrometry; and Aim II. To initiate a functional analysis of the PfVPA and PfVPR proteins in P. falciparum. PUBLIC HEALTH RELEVANCE: Malaria parasites invade and ultimately destroy circulating red blood cells of their host, often leading to severe clinical illness or death. During their intraerythrocytic development, malaria parasites switch expression to alternative forms of a variant surface protein PfEMP1 encoded by the var genes to evade the host immune response. The goal of this research collaboration is to combine tools and techniques from genetics, molecular biology, and immunochemistry, and the expertise of two laboratories to unravel the molecular determinants of antigenic variation during malaria infection.

描述（由申请人提供）：本研究项目的目标是开发一种结合染色质免疫沉淀、串联亲和纯化和质谱的新方法，以鉴定编码疟疾毒力因子、恶性疟原虫红细胞膜蛋白1 （PfEMP1）的var多基因家族可能的激活因子和/或抑制因子。在它们的红细胞发育过程中，疟疾寄生虫转换表达这种变体表面蛋白的替代形式以逃避宿主的免疫反应。每个恶性疟原虫的基因组包含大约60个变异基因，分布在不同的染色体上。然而，每个寄生虫只有一个var基因以互斥的方式表达。var基因的启动子区域在这一调控中起着重要作用，据信这一机制可能涉及特异性抑制因子（PfVPR）和/或激活因子（PfVPA）分子。本提案的具体目标包括实验设计和方法，其中一些是我们实验室新开发的恶性疟原虫的串联亲和纯化和荧光原位杂交（FISH）。目的：利用染色质免疫沉淀、串联亲和纯化和质谱技术鉴定PfVPA和PfVPR基因表达调控因子；和Aim II。启动恶性疟原虫PfVPA和PfVPR蛋白的功能分析。