COBRE: NDSU: PROJECT 5: NEW INVESTIGATOR

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 A节：具体目标 我实验室研究的最终目标是使用各种结构和生化 理解自噬等途径中蛋白质功能的原子基础的技术 (一种降解细胞内物质的溶酶体靶向途径)与先天免疫 途径；这些途径之间的串扰；这些途径/蛋白质在 对病原体的防御；以及不同病原体对这些蛋白的抑制。一 该项目重点研究Beclin1的结构、功能和分子机制， 一种必不可少的自噬效应器。我们之前的工作，部分资金来自NIH R21拨款， 探讨细胞和病毒Bcl2蛋白抑制小梁形成的分子机制 1和自噬。我们计划继续我们对Beclin 1蛋白的研究，包括其 与其他自噬蛋白和病原体编码蛋白相互作用。另一个 该项目的重点是RIG-I，这是抗病毒天然免疫的关键介体。我们计划调查 RIG-I在先天性防御中作用的原子基础通过研究分子内、内和外 传递信号的RIG-I卡域的通路或交叉通路相互作用 触发或抑制各种通路。 所有这些研究的前提是获得纯的、同质的、重组的 蛋白。我们正在使用Cobre基金来生产这些试剂并获得初步数据 申请其他拨款以进一步研究这些途径。眼镜蛇的具体目标 资金用于大量超量表达和纯化下列重组蛋白 适用于后续的结构和生化研究： +目标1：不同的Beclin 1结构域。 +目标2：RIG-I卡域。 +AIM 3：其下游信令合作伙伴IPS-1的卡域。 +Aim 4：RIG-I和LGP2(RIG-I同源物)的抑制因子结构域。 +Aim 5：与RIG-I和IPS-1相互作用的基本自噬效应因子ATG5 卡片域。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Section A: Specific Aims The ultimate goal of research in my laboratory is to use various structural and biochemical techniques to understand the atomic basis of function of proteins of pathways like autophagy (a lysosomal targeting pathway for degradation of intracellular objects) and innate immune pathway; the cross-talk between these pathways; the role of these pathways/proteins in the defense against pathogens; and the inhibition of these proteins by different pathogens. One project focuses on investigating the structure, function and molecular mechanism of Beclin1, an essential autophagy effector. Our previous work, funded in part by an NIH R21 grant, investigated the molecular mechanism by which cellular and viral Bcl-2 proteins inhibit Beclin 1 and autophagy. We plan to continue our investigation of the Beclin 1 protein, including its interaction with other autophagy proteins and with pathogen-encoded proteins. Another project focuses on Rig-I, a key mediator of anti-viral innate immunity. We plan to investigate the atomic basis of the role of Rig-I in innate defenses by studying the intra-molecular, intra- pathway or cross-pathway interactions of the Rig-I CARD domains that transmit signals to trigger or repress various pathways. A prerequisite of all these studies is the availability of pure, homogeneous, recombinant protein. We are using COBRE funding to produce these reagents and obtain preliminary data to apply for other grants to further investigate these pathways. Specific aims of the COBRE funding are to overexpress and purify the recombinant proteins listed below, in quantities suitable for subsequent structural and biochemical studies: + Aim 1: Different Beclin 1 domains. + Aim 2: Rig-I CARD domains. + Aim 3: CARD domains of IPS-1, its downstream signaling partner. + Aim 4: Repressor domains of Rig-I and LGP2 (a Rig-I homolog). + Aim 5: The essential autophagy effector Atg5, which interacts with the Rig-I and IPS-1 CARD domains.