The Apical Complex: a Targeted Investigation of the Molecular Functions of this Structure Essential to Apicomplexan Parasite Invasion and Replication.

顶端复合体：对顶端复合体寄生虫入侵和复制所必需的该结构的分子功能进行有针对性的研究。

• 批准号: MR/M011690/1
• 负责人: Ross Waller
• 金额: $ 57.44万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FM011690%2F1
• 关键词: Apical; Complex; Targeted; Investigation; Molecular

Apicomplexans are a large group of single-celled pathogens that cause significant disease in humans. The most notorious member of this group causes malaria, a devastating disease in many developing regions of the world and responsible for over 500 million cases, and 0.6 million deaths per year. Other apicomplexan species cause widespread human diseases: cryptosporidiosis, a leading cause of fatal infant diarrhoea; and toxoplasmosis, which infects approximately 30% of all humans. Further, several apicomplexans infected domesticated animals causing significant loss to human food production.All apicomplexans cause disease by growing and dividing within the cells of their human (or animal) hosts. They enter their host's cells by non-destructively forming a temporary pore in the host cell wall, through which they slide in and then reseal. By keeping the host cell alive it provides a constant source of food and nutrients to the parasite. Only once the parasites have divided to many times the original number do they burst open and kill the human cell, and then actively seek out and invade new host cells.The key structure used by the pathogen during invasion of the host's cells is a structure called the apical complex, and this is a shared feature of all apicomplexan pathogens. The apical complex can be observed using microscopes as an internally reinforced point at the tip of the cell that acts as a cellular nozzle or syringe. At its apex is a small opening, through which it first releases sticky molecules onto the pathogens surface that help it adhere to its host's cells. Subsequently, it uses this opening to inject invasion factors directly into its host that create the invasion pore through which it enters the host. Further pathogen molecules are then released from the apical complex that enable it to feed on its host. Thus, the apical complex is at the heart of these coordinated events of host cell invasion, and is therefore an essential structure for disease formation.Despite the importance of the apical complex, this structure remains poorly understood. Few of the molecular components are known, and this limits understanding of its architecture, assembled, and how it provides the general functions that it does. This study will use multiple approaches to identify and catalogue the molecular components of the apical complex. It will employ the apicomplexan Toxoplasma as the best developed experimental system for apicomplexans. A detailed model of the architecture of the apical complex will be generated, and using genetic tagging of individual structural components, their assembly and behaviour will be characterised in live cells, including during the invasion events. The function of the individual components, and of the apical complex as a whole, will then be dissected by selectively removing one component at a time at a genetic level. This will provide a mechanistic understanding of how the apical complex achieves the deadly tasks of host cell penetration and exploitation.An integrated understanding of the apical complex organisation and function will provide clearer insights into how apicomplexan organisms have achieved such success as pathogens. Moreover, understanding their mechanisms of pathogenesis provides the best opportunity for strategically designing disease treatment strategies.

顶复门是一大群单细胞病原体，在人类中引起重大疾病。这一组中最臭名昭著的成员引起疟疾，这是世界上许多发展中地区的一种毁灭性疾病，每年造成5亿多病例和60万人死亡。其他apicomplexan物种引起广泛的人类疾病：隐孢子虫病，致命的婴儿腹泻的主要原因;弓形虫病，感染约30%的人类。此外，一些顶复门感染家养动物，导致人类粮食生产的重大损失。所有顶复门都通过在其人类（或动物）宿主的细胞内生长和分裂而引起疾病。它们通过非破坏性地在宿主细胞壁上形成一个临时孔进入宿主细胞，通过这个孔它们滑进去，然后重新密封。通过保持宿主细胞存活，它为寄生虫提供了恒定的食物和营养来源。只有当寄生虫分裂到原来数量的许多倍时，它们才会突然打开并杀死人体细胞，然后积极寻找并入侵新的宿主细胞。病原体在入侵宿主细胞时使用的关键结构是一种称为顶端复合体的结构，这是所有顶端复合体病原体的共同特征。顶端复合体可以用显微镜观察到，作为细胞尖端的内部加强点，充当细胞喷嘴或注射器。在它的顶端是一个小开口，它首先通过这个开口将粘性分子释放到病原体表面，帮助它粘附在宿主细胞上。随后，它利用这个开口将侵入因子直接注入其宿主，从而产生侵入孔，通过该孔进入宿主。然后从顶端复合体释放出更多的病原体分子，使其能够以宿主为食。因此，顶端复合体是这些宿主细胞入侵的协调事件的核心，因此是疾病形成的必要结构。尽管顶端复合体的重要性，这种结构仍然知之甚少。很少有分子组分是已知的，这限制了对其结构、组装以及如何提供其一般功能的理解。本研究将使用多种方法来识别和编目顶端复合物的分子组分。它将采用顶复门弓形虫作为顶复门的最佳开发的实验系统。将生成顶端复合体结构的详细模型，并使用单个结构组件的遗传标记，它们的组装和行为将在活细胞中进行表征，包括在入侵事件期间。然后，通过在遗传水平上一次选择性地去除一个组分，将单个组分的功能以及顶端复合体作为一个整体的功能进行解剖。这将提供一个机械的理解，顶端复合体如何实现致命的任务，宿主细胞的渗透和exploitation.A顶端复合体的组织和功能的综合理解将提供更清晰的见解apicomplexan生物如何取得了这样的成功，病原体。此外，了解其发病机制为战略性设计疾病治疗策略提供了最佳机会。

项目成果

A Comprehensive Subcellular Atlas of the Toxoplasma Proteome via hyperLOPIT Provides Spatial Context for Protein Functions.

• DOI: 10.1016/j.chom.2020.09.011
• 发表时间: 2020-11-11
• 期刊: Cell host & microbe
• 影响因子: 30.3
• 作者: Barylyuk K;Koreny L;Ke H;Butterworth S;Crook OM;Lassadi I;Gupta V;Tromer E;Mourier T;Stevens TJ;Breckels LM;Pain A;Lilley KS;Waller RF
• 通讯作者: Waller RF

Two essential Thioredoxins mediate apicoplast biogenesis, protein import, and gene expression in Toxoplasma gondii.

• DOI: 10.1371/journal.ppat.1006836
• 发表时间: 2018-03
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Biddau M;Bouchut A;Major J;Saveria T;Tottey J;Oka O;van-Lith M;Jennings KE;Ovciarikova J;DeRocher A;Striepen B;Waller RF;Parsons M;Sheiner L
• 通讯作者: Sheiner L

A Pentatricopeptide Repeat Protein in the Plasmodium apicoplast is essential and shows sequence-specific RNA binding

疟原虫顶质体中的五肽重复蛋白是必需的，并且显示出序列特异性 RNA 结合

• DOI: 10.1101/388728
• 发表时间: 2018
• 作者: Hicks J