Structure-function relationships of Chlamydia trachomatis Tarp & binding proteins

沙眼衣原体 Tarp 的结构与功能关系

• 批准号: 7638033
• 负责人: Travis James Jewett
• 金额: $ 15.53万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7638033
• 关键词: 1-Phosphatidylinositol 3-Kinase; Actins; Antibiotics; Applications Grants; Bacteria; Bacterial Proteins; Binding; Binding Proteins; Biological Assay; Blindness; Case Study; Cell physiology; Cells; Chemicals; Chimeric Proteins; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Clinical; Data; Defect; Development; Disease; Drug Delivery Systems; Event; Genome; Goals; Human; Immunofluorescence Immunologic; Immunoprecipitation; Infection; Lipids; Maps; Modification; Molecular; Morbidity - disease rate; Pathogenesis; Pathway interactions; Phosphorylation; Phosphotransferases; Play; Principal Investigator; Proline; Proteins; Publishing; Receptor Protein-Tyrosine Kinases; Recombinants; Recruitment Activity; Research Proposals; Role; Sexually Transmitted Diseases; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Src homology 2 domain-containing, transforming protein 1; Staging; Structure-Activity Relationship; Syringes; Tertiary Protein Structure; Type III Secretion System Pathway; Tyrosine; Tyrosine Phosphorylation Site; Vaccines; Virulence Factors; design; inhibitor/antagonist; novel; novel therapeutic intervention; pathogen; protein protein interaction; residence; resistant strain; src Homology Domains

DESCRIPTION (provided by applicant): The obligate intracellular bacterium Chlamydia trachomatis causes substantial morbidity in the US and world wide. An inability to manipulate the chlamydial genome has consequently produced a dearth of bona fide virulence factors. Hence, pathogenic mechanisms of chlamydial infections are poorly described and require further examination. A C. trachomatis effector called Tarp, for translocated actin recruiting protein is a candidate virulence factor. Tarp is tyrosine phosphorylated by a host cell kinase and is associated with actin recruitment during Chlamydiae entry. Tarp protein domains have been identified and include: i) an actin binding and nucleating domain, ii) a proline rich oligomerization domain and iii) a phosphorylation domain. The function of the Tarp phosphorylation domain is currently unknown and the focus of the attached grant proposal. Src-homology domain 2 (SH2) containing proteins are candidate Tarp interacting proteins. The purpose of this grant proposal is to identify Tarp binding proteins (those with and without SH2 domains) by using assays to detect protein-protein interactions such as GST-fusion pull-down assays, immunoprecipitation, and two commercially available assays designed to target SH2 domain containing proteins. And secondly, to assess any requirements for identified Tarp binding proteins in Chlamydiae entry and development. Recently, PIS kinase was implicated in association with phosphorylated Tarp. Modification of lipids by PI3K ultimately regulates a variety of cellular processes. We predict that Tarp recruitment of PI3K plays a vital role in the establishment of chlamydial infection and intend to examine a requirement for this association with chemical inhibits and interfering RNAs. Currently, a chlamydial vaccine is not available. Elucidation of Tarp interactions with the host is crucial for mapping signaling pathways used by C. trachomatis to establish residence within infected cells. A better understanding of the mechanisms employed by Chlamydiae to initiate a successful infection will result in the identification of novel therapeutic interventions. RELEVANCE: The sexually transmitted disease causing bacteria, Chlamydia trachomatis, infected more than 1 million people in the US alone last year. A recently identified Chlamydiae protein called Tarp is believed to play an important role in the initial stages of infection. The goal of this research proposal is to determine how the Tarp protein facilitates Chlamydiae to cause disease.

描述(由申请人提供)：专性细胞内沙眼衣原体细菌在美国和世界范围内导致大量发病率。因此，由于无法操纵衣原体基因组，因此缺乏真正的毒力因子。因此，衣原体感染的致病机制尚未得到很好的描述，需要进一步研究。沙眼衣原体的一种名为Tarp的效应器，即易位肌动蛋白招募蛋白，是一个候选的毒力因子。TARP是被宿主细胞激酶磷酸化的酪氨酸，与衣原体进入时的肌动蛋白募集有关。TARP蛋白结构域已被确定，包括：i)肌动蛋白结合和核化区，ii)富含脯氨酸的寡聚区和iii)磷酸化结构域。TARP磷酸化结构域的功能目前尚不清楚，也是所附赠款提案的重点。含有SRC同源结构域2(SH2)的蛋白是候选的Tarp相互作用蛋白。这项拨款提案的目的是通过使用检测蛋白质-蛋白质相互作用的方法来鉴定Tarp结合蛋白(那些有和没有SH2结构域的蛋白质)，例如GST融合下拉试验、免疫沉淀和两种针对含有SH2结构域的蛋白质的商业可用的分析方法。其次，评估衣原体进入和发育过程中对已鉴定的TARP结合蛋白的任何需求。最近，PIS激酶被认为与磷酸化的TARP有关。PI3K对脂类的修饰最终调节了多种细胞过程。我们预测PI3K的Tarp募集在衣原体感染的建立中起着至关重要的作用，并打算研究这种联系与化学抑制和干扰RNA的要求。目前，尚无衣原体疫苗可用。阐明TARP与宿主的相互作用对于定位沙眼衣原体在感染细胞内驻留的信号通路至关重要。更好地了解衣原体启动成功感染的机制将导致新的治疗干预措施的确定。 相关性：引起细菌的性传播疾病沙眼衣原体去年仅在美国就感染了100多万人。最近发现的一种名为Tarp的衣原体蛋白被认为在感染的初始阶段发挥了重要作用。这项研究提案的目标是确定Tarp蛋白如何促进衣原体致病。