Global mechanisms for control of the trypanosome proteome: Defining the composition, origins and roles of cullin E3 ligases.

控制锥虫蛋白质组的全局机制：定义 cullin E3 连接酶的组成、起源和作用。

• 批准号: MR/P009018/1
• 负责人: Mark Field
• 金额: $ 43.88万
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FP009018%2F1
• 关键词: Global; mechanisms; control; trypanosome; proteome

Parasitic protozoa are major agents of disease, and afflict a major proportion of the global population. In evolutionary terms parasites are highly removed from their hosts, and this is frequently reflected in the presence of unique or unusual mechanisms that underpin their biology. This aspect presents multiple opportunities for the understanding of pathogenesis, the possibility of identifying therapeutic targets as well as offering a fascinating evolutionary perspective on many cellular processes. African trypanosomes are totally reliant on antigenic variation for their longterm survival. The process is so successful that trypanosomes can infect a mammalian host for many months and in some cases years/decades. Additional immune evasion mechanisms, which defend the parasite against components of both the innate and acquired arms of the host immune response, require surface or endosome-located proteins. We have recently made three significant advances in understanding the cell biology that underpins these immune evasion processes; 1. Characterisation of the cell surface proteome, which revealed both remarkable diversity from higher eukaryotes and even from other trypanosomatids such as American trypanosomes, 2. Demonstration of partitioning of surface components into at least four distinct micro-domains - surface, flagellar pocket, flagellum and endosome, with some proteins exhibiting a combination between all four, and 3. Identifying components of the ubiquitylation machinery that control the expression levels of surface proteins. These new insights open a route to understanding how the unique pathogenic surface of African trypanosomes is regulated at the molecular level, together with dissecting how such regulation contributes towards basic biology and pathogenesis. Coupled with this is the means to exploit proteomics technology and insights developed recently to understand in a more global manner regulation of the trypanosome cellular proteome. We propose to interrogate in detail how a group of trypanosome ubiquitin ligases, called cullins, function in this context. Ubiquitin is a major mediator of protein turnover pathways, with the cullin group especially important due to roles in surface receptor turnover and modulation of cell cycle progression. Given the amenability of the trypanosome for genetic manipulation, the comparatively small size of the proteome and the emphasis on post-transcriptional and translation-linked mechanisms to control protein expression levels, we propose that the trypanosome provides a very attractive approach to understand these ubiquitylating complexes, their functional stratification and specificity, in addition to how they contribute towards trypanosome biology and infectivity.

寄生原生动物是疾病的主要媒介，并且折磨全球人口的主要部分。从进化的角度来看，寄生虫与宿主的距离很远，这经常反映在支撑其生物学的独特或不寻常的机制上。这一方面为理解发病机制提供了多种机会，确定治疗靶点的可能性，并为许多细胞过程提供了迷人的进化视角。 非洲锥虫完全依赖于抗原变异来长期生存。这个过程是如此成功，锥虫可以感染哺乳动物宿主数月，在某些情况下几年/几十年。额外的免疫逃避机制，保护寄生虫免受宿主免疫应答的先天性和获得性臂的成分，需要表面或内体定位的蛋白质。我们最近在理解支持这些免疫逃避过程的细胞生物学方面取得了三项重大进展; 1.细胞表面蛋白质组的特征，揭示了显着的多样性，从高等真核生物，甚至从其他锥虫，如美国锥虫，2。证明表面组分分成至少四个不同的微域-表面、鞭毛口袋、鞭毛和内体，其中一些蛋白质表现出所有四个之间的组合，并且3。确定控制表面蛋白表达水平的泛素化机制的组成部分。 这些新的见解开辟了一条途径，以了解非洲锥虫独特的致病表面如何在分子水平上进行调节，以及解剖这种调节如何有助于基础生物学和发病机制。与此相结合的是利用蛋白质组学技术的手段和最近开发的见解，以更全面的方式了解锥虫细胞蛋白质组的调控。我们建议详细询问一组锥虫泛素连接酶，称为cullins，在这种情况下的功能。泛素是蛋白质周转途径的主要介质，其中cullin基团由于在表面受体周转和调节细胞周期进程中的作用而特别重要。鉴于顺从的锥虫的遗传操作，相对较小的大小的蛋白质组和强调转录后和抑制相关的机制来控制蛋白质的表达水平，我们建议锥虫提供了一个非常有吸引力的方法来了解这些泛素化复合物，其功能分层和特异性，除了它们如何有助于锥虫生物学和感染性。

项目成果

Evolution of cullin E3 ubiquitin ligases and function in trypanosomes

• DOI: 10.1101/2023.07.24.550360
• 发表时间: 2023-07
• 期刊: bioRxiv
• 作者: R. C. del Pino;M. Zoltner;Kayo Yamada;Erin R Butterfield;Mark C. Field

Pore timing: the evolutionary origins of the nucleus and nuclear pore complex.

• DOI: 10.12688/f1000research.16402.1
• 发表时间: 2019-01-01
• 期刊: F1000Research
• 作者: Field, Mark C;Rout, Michael P
• 通讯作者: Rout, Michael P

Evolution and diversification of the nuclear envelope.

• DOI: 10.1080/19491034.2021.1874135
• 发表时间: 2021-12
• 期刊: Nucleus (Austin, Tex.)
• 作者: Padilla-Mejia NE;Makarov AA;Barlow LD;Butterfield ER;Field MC
• 通讯作者: Field MC

A leucine aminopeptidase is involved in kinetoplast DNA segregation in Trypanosoma brucei.

• DOI: 10.1371/journal.ppat.1006310
• 发表时间: 2017-04
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Peña-Diaz P;Vancová M;Resl C;Field MC;Lukeš J
• 通讯作者: Lukeš J

Impact of inherent biases built into proteomic techniques: Proximity labeling and affinity capture compared.

蛋白质组学技术内置的固有偏见的影响：比较接近性标记和亲和力捕获。

• DOI: 10.1016/j.jbc.2022.102726
• 发表时间: 2023-01
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Moreira, Claudia Maria do Nascimento;Kelemen, Cristina D.;Obado, Samson O.;Zahedifard, Farnaz;Zhang, Ning;Holetz, Fabiola B.;Gauglitz, Laura;Dallagiovanna, Bruno;Field, Mark C.;Kramer, Susanne;Zoltner, Martin
• 通讯作者: Zoltner, Martin