Systematic analysis of cell death regulation in mosquitoes

标题：蚊子细胞死亡规律的系统分析。 细胞死亡(细胞凋亡)是载体/宿主免疫反应的重要组成部分 机制。它与多种病原体的感染生命周期有关，如 蚊子体内的疟疾寄生虫和西尼罗河病毒。然而，我们对细胞死亡的理解 由于缺乏一份完整的主要昆虫名单，对蚊子的监管受到了极大的阻碍 细胞死亡调节剂，即*Reaper/HID/Grim-like IAP拮抗剂，在注释蚊子中 基因组。 IAP(细胞凋亡抑制蛋白)是核心细胞死亡机制的重要刹车 在昆虫身上。它们与半胱氨酸天冬氨酸酶结合，抑制这些致命酶的活性。遗传学研究 在果蝇中的研究表明，发育过程中大多数程序性细胞死亡是通过 IAP拮抗剂如REPAR、HID和GRIM(RHG)在注定要死亡的细胞中的特异性表达。 这些IAP拮抗剂中的大多数也是调节细胞死亡以响应环境的需要。 压力。疟疾主要媒介冈比亚按蚊的基因组计划揭示了一种 与果蝇相比，IAP和caspase基因家族显著扩大。这是 假设反映了微调细胞死亡调节的功能要求，以响应 寄生虫和病毒感染。这也强烈地暗示了IAP拮抗剂？？休息室？？半胱氨酸酶 与果蝇一样，途径对于蚊子细胞死亡的调控是至关重要的。 然而，由于广泛的序列差异，注释的完整的An.冈比亚亚纲基因组 序列最初不包括任何类似于RHG的IAP-拮抗剂的同源基因。使用系统化的 利用生物信息学的方法，我们鉴定并鉴定了第一批类重组人凝血酶原激活蛋白拮抗剂， Michelob_x(MX)，在按蚊和伊蚊基因组中。为了调查这些人的参与 在媒介-病原体相互作用中的促凋亡基因，我们在库蚊中鉴定了MX同源基因 并发现它是由库那普病毒(一种蚊虫杆状病毒)感染奎纳乌氏库蚊而诱导的 (第4.5节；周等人，正在准备中)。此外，我们还发现了另一种IAP拮抗剂， IBMP6，作为果蝇HID的同源基因。有趣的是，ibmp6是由MIV感染的 Aag2细胞(4.6节)。这些数据表明，与果蝇一样，多个IAP拮抗剂是 蚊子细胞死亡调控所必需的。此外，还使用了不同的IAP拮抗剂 对抗不同的感染或在不同的组织中。 由于iap拮抗剂基因在昆虫细胞死亡调控中起着关键作用，因此我们的工作 揭示了它们参与了病原体诱导的媒介反应，充分阐明了这组 蚊子中的基因将是全面了解细胞死亡调控的关键 对病原体攻击的反应以及在正常蚊子生命周期中。这项拟议的项目 目的：(1)大剂量IAP拮抗剂的系统鉴定和功能验证 蚊子基因组。(2)阐明这些IAP拮抗剂在媒介病原体中的作用 利用CuniNPV、MIV和B.algerae进行互作。(三)系统开发公共资源 蚊子细胞死亡规律的分析。 细胞死亡(细胞凋亡)与多种病原体的感染生命周期有关，如疟疾 蚊子体内的寄生虫和西尼罗河病毒。了解细胞死亡反应将有助于我们发展 控制蚊子传播疾病和控制蚊子数量的战略。

Title: Systematic analysis of cell death regulation in mosquitoes. Cell death (apoptosis) is an important component of the vector/host immuno-response mechanism. It has been implicated in the infectious life cycle of a variety of pathogens, such as the malaria parasite and the West Nile virus, in mosquitoes. However, our understanding of cell death regulation in mosquitoes has been hindered greatly by the lacking of a complete list of major insect cell death regulators, i.e. the *Reaper/Hid/Grim -like IAP-antagonists, in the annotated mosquito genomes. IAPs (Inhibitor of Apoptosis Proteins) are the essential ¿Brake¿ of the core cell death machinery in insects. They bind to caspases and inhibit the activity of these ¿deadly¿ enzymes. Genetic studies in Drosophila have shown that most programmed cell death during development is mediated through specific expression of IAP-antagonists such as reaper, hid, and grim (RHG) in cells destined to die. Most of these IAP-antagonists are also required for mediating cell death in response to environmental stress. The genome project for the primary malaria vector, Anopheles gambiae, revealed a significant expansion of the IAP and caspase gene families when compared to Drosophila. This was postulated to reflect the functional requirement of fine-tuning cell death regulation in response to parasite and virus infection. It also strongly implies that the IAP-antagonist ?? IAP ?? Caspases pathway, as is in Drosophila, is fundamentally important for cell death regulation in mosquitoes. However, due to extensive sequence divergence, the annotated complete An. gambiae genome sequence did not initially include any ortholog of RHG-like IAP-antagonist. Using a systematic bioinformatics approach, we identified and subsequently characterized the first RHG-like IAPantagonists, michelob_x (mx), in Anopheles and Aedes genomes. To probe the involvement of these pro-apoptotic genes in mediating vector-pathogen interaction, we identified the mx ortholog in Culex and found that it is induced by CuniNPV (a mosquito baculovirus) infection to Cx.quinauifasciatus (section 4.5; Zhou et al, in preparation). In addition, we have identified another IAP-antagonist, IBMP6, as the ortholog of Drosophila Hid. Interestingly, ibmp6 is induced by MIV infection of the Aag2 cells (section 4.6). These data indicate that like in Drosophila, multiple IAP-antagonist are required for cell death regulation in mosquito. Further more, distinct IAP-antagonists is(are) utilized against different infections or in different tissues. Since the IAP-antagonist genes play pivotal roles in insect cell death regulation and our work revealed that they are involved in pathogen-induced vector response, elucidating fully this group of genes in mosquitoes will be essential for the comprehensive understanding of cell death regulation in response to pathogen challenge as well as during normal mosquito life cycle. This proposed project aims at (1) Systematic identification and functional verification of the IAP-antagonists in major mosquito genomes. (2)Elucidating the role of these IAP-antagonists in mediating vector-pathogen interaction using CuniNPV, MIV, and B.algerae. (3) Developing public resources for systematic analysis of cell death regulation in mosquitoes. Cell death (apoptosis) is implicated in the infectious life cycle of a variety of pathogens, such as the malaria parasite and the West Nile virus, in mosquitoes. Understanding cell death response will help us to develop strategy for controlling mosquito transmitted disease and for controlling mosquito population.