MT VET COBRE PROJECT 1: HOST-PATHOGEN COMMUNICATION IN TOXOPLASMA

MT VET COBRE 项目 1：弓形虫中的宿主-病原体通讯

• 批准号: 7960525
• 负责人: JAY R RADKE
• 金额: $ 14.53万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960525
• 关键词: Automobile Driving; Cells; Centers of Research Excellence; Chronic; Communication; Computer Retrieval of Information on Scientific Projects Database; Development; Emerging Communicable Diseases; Funding; Gene Transfer; Genes; Grant; Hela Cells; Histones; Infection; Institution; Lysine; Maps; Methylation; Molecular; Mutation; Parasites; Pathogenesis; Proteins; Relative (related person); Research; Research Personnel; Resistance; Resources; SET Domain; Site-Directed Mutagenesis; Source; Testing; Toxoplasma; United States National Institutes of Health; Work; pathogen; resistant strain

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Molecular mechanisms controlling bradyzoite differentiation in the parasite are completely unknown. Dr. Radke has shown previously that Toxoplasma Type I strains are naturally resistant to the artificially induced bradyzoite differentiation and to over-expression of some host cell genes. Dr. Radke's lab uses (1) a complementation strategy to transfer a gene from the Type I RH strain to a Type III strain CTG parasite to identify genes that may render Type I strains resistant to development; and (2) testing histone methylation as one primary mechanism driving induced parasite development when distinct host cell genes are over-expressed in the infected cell. They have transferred a gene from the Type I RH strain to a Type III strain CTG parasite and recovered sequence that maps to ChrVIII beginning at 6300 Kbp and ChrXII at 1200 Kbp. Present work will focus on narrowing the candidate resistance sequences to a single gene that may underlie the molecular mechanisms controlling bradyzoite differentiation and the pathogenesis of chronic infection in the host. Dr. Radke has recently shown that new methylation at lysines 4, 9, and 36 of histone 3 occur in concert with the over-expression of CDA1 in HeLa cells, but whether this is a direct action of CDA1 is unknown. Work to confirm (or not) direct methylation of core histone residues by CDA1 will be completed using site-directed mutagenesis. Mutation of select residues in the SET domain that are able to reduce methylation at lysine 4, 9, 27, or 36 relative to the wild-type protein would suggest CDA1 is able to directly methylate histone residues.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 控制寄生虫中缓殖子分化的分子机制完全未知。 Radke博士先前已经表明，弓形虫I型菌株对人工诱导的缓殖子分化和某些宿主细胞基因的过度表达具有天然抗性。 Radke博士的实验室使用（1）互补策略将基因从I型RH菌株转移到III型菌株CTG寄生虫，以鉴定可能使I型菌株对发育具有抗性的基因;和（2）当不同的宿主细胞基因在感染细胞中过度表达时，测试组蛋白甲基化作为驱动诱导寄生虫发育的主要机制。 他们将一个基因从I型RH株转移到III型CTG株寄生虫中，并恢复了从6300 Kbp开始映射到ChrVIII和从1200 Kbp开始映射到ChrXII的序列。 目前的工作将集中在缩小候选耐药序列的一个单一的基因，可能是控制缓殖子分化和慢性感染的发病机制在主机的分子机制。 Radke博士最近表明，组蛋白3的赖氨酸4、9和36处的新甲基化与HeLa细胞中CDA1的过度表达一致，但这是否是CDA1的直接作用尚不清楚。 将使用定点诱变完成确认（或不确认）核心组蛋白残基被CDA1直接甲基化的工作。 相对于野生型蛋白，SET结构域中能够减少赖氨酸4、9、27或36处的甲基化的选择残基的突变将表明CDA1能够直接甲基化组蛋白残基。