Elucidating the Cep135 - CPAP- STIL protein interaction network behind primary microcephaly and centriole formation

阐明原发性小头畸形和中心粒形成背后的 Cep135 - CPAP-STIL 蛋白相互作用网络

• 批准号: MR/N009274/1
• 负责人: Ioannis Vakonakis
• 金额: $ 54.03万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FN009274%2F1
• 关键词: Elucidating; Cep135; CPAP; STIL; protein

Primary microcephaly is a hereditary disease characterised by reduced brain size from birth and mental retardation. It occurs in ~1 in 10,000 individuals in some populations, but, importantly, it is one of few diseases where we can directly trace the effects of single mutations to brain development and cognitive functions. As a result primary microcephaly cases have proved instructive in identifying crucial brain development factors for which no backup systems exist.Centrosomes, are small organelles in human cells that organise a network of thin filaments (known as microtubules) that are essential for cells to grow, duplicate, move and sense their surroundings. Defects in centrosomes have been implicated in microcephaly. Five out of nine genes known to cause the disease correspond to centrosome components, and these include three proteins known to be essential for the formation of these organelles. In addition to microcephaly, centrosomal defects are causative agents for multiple human medical conditions, including male sterility, ciliopathies and possibly cancer. Thus, understanding how centrosomes form is an important biological question with direct medical relevance.Over the last few years our group, and others, have shown how a single protein, SAS-6, forms the initial framework onto which centrosomes are build. Crucial to this understanding was a combination of biophysical, structural and cell biology tools that allowed us to analyse the shape of essential proteins, envision how such proteins might join to form molecular machines, and test these insights in human cells. Here, we propose to build upon our understanding of the initial centrosomal framework by studying three protein components (Cep135, CPAP and STIL) that link to it. We have selected these components because they are essential for centrosomes, they appear to be connected to one another and to SAS-6, and importantly, they are all directly implicated in primary microcephaly. We believe that understanding the role of these three proteins will also inform us on how centrosomes are formed in normal cells and how defects in them cause severe diseases. We expect that these results will underpin future efforts on how to treat such diseases.Our group has long experience in the biophysical and structural biology methods necessary for the pursuit of this project. However, we do not rely on our core competencies alone. Our goal of understanding the centrosome structure is shared with internationally acclaimed groups in Oxford and abroad, with whom we collaborate. Our network of laboratories provides the broadest possible base of technical expertise and, thus, the best hope for determining how centrosomes form.

原发性小头畸形是一种遗传性疾病，其特征是出生时大脑体积缩小和智力迟钝。在某些人群中，它发生在1/10，000的个体中，但重要的是，它是少数几种我们可以直接追踪单一突变对大脑发育和认知功能影响的疾病之一。因此，原发性小头畸形病例已被证明在确定关键的大脑发育因素方面具有指导意义，而这些因素并不存在备份系统。中心体是人类细胞中的小细胞器，它组织一个细丝网络（称为微管），对细胞生长，复制，移动和感知周围环境至关重要。中心体缺陷与小头畸形有关。已知引起这种疾病的九个基因中有五个与中心体成分相对应，其中包括三种已知对这些细胞器的形成至关重要的蛋白质。除了小头畸形，中心体缺陷也是多种人类疾病的致病因素，包括男性不育、纤毛病和可能的癌症。因此，了解中心体是如何形成的是一个重要的生物学问题，具有直接的医学相关性。在过去的几年里，我们的小组和其他人已经展示了一个单一的蛋白质SAS-6是如何形成中心体的初始框架的。对这种理解至关重要的是生物物理，结构和细胞生物学工具的结合，使我们能够分析必需蛋白质的形状，设想这些蛋白质如何结合形成分子机器，并在人类细胞中测试这些见解。在这里，我们建议建立在我们的理解的初始中心体框架，通过研究三个蛋白质组分（Cep 135，CPAP和STIL），链接到它。我们选择这些组件，因为它们是必不可少的中心体，他们似乎是连接到另一个和SAS-6，重要的是，他们都直接牵连到原发性小头畸形。我们相信，了解这三种蛋白质的作用也将告诉我们中心体如何在正常细胞中形成，以及它们的缺陷如何导致严重的疾病。我们希望这些结果将为未来治疗此类疾病的努力奠定基础。我们的团队在追求该项目所需的生物物理学和结构生物学方法方面拥有长期经验。然而，我们并不单单依靠我们的核心能力。我们理解中心体结构的目标是与牛津和国外的国际知名团体分享，我们与他们合作。我们的实验室网络提供了最广泛的技术专业知识基础，因此，最有希望确定中心体如何形成。

项目成果

How to Break a Ring: Exploring the Mechanisms of SAS-6 Oligomerisation

如何打破环：探索 SAS-6 寡聚化机制

• DOI: 10.1016/j.bpj.2016.11.3047
• 发表时间: 2017
• 期刊: Biophysical Journal
• 影响因子: 3.4
• 作者: Busch J

Interaction between the Caenorhabditis elegans centriolar protein SAS-5 and microtubules facilitates organelle assembly.

• DOI: 10.1091/mbc.e17-06-0412
• 发表时间: 2018-03-15
• 期刊: Molecular biology of the cell
• 影响因子: 3.3
• 作者: Bianchi S;Rogala KB;Dynes NJ;Hilbert M;Leidel SA;Steinmetz MO;Gönczy P;Vakonakis I
• 通讯作者: Vakonakis I

The 3-phosphoinositide-dependent protein kinase 1 is an essential upstream activator of protein kinase A in malaria parasites.

• DOI: 10.1371/journal.pbio.3001483
• 发表时间: 2021-12
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Hitz E;Wiedemar N;Passecker A;Graça BAS;Scheurer C;Wittlin S;Brancucci NMB;Vakonakis I;Mäser P;Voss TS
• 通讯作者: Voss TS

Structural analysis of P. falciparum KAHRP and PfEMP1 complexes with host erythrocyte spectrin suggests a model for cytoadherent knob protrusions.

恶性疟原虫KAHRP和PFEMP1复合物与宿主红细胞光谱蛋白的结构分析提出了一个细胞辅助旋钮突起的模型。

• DOI: 10.1371/journal.ppat.1006552
• 发表时间: 2017-08
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Cutts EE;Laasch N;Reiter DM;Trenker R;Slater LM;Stansfeld PJ;Vakonakis I
• 通讯作者: Vakonakis I

Structures of SAS-6 coiled coil hold implications for the polarity of the centriolar cartwheel.

• DOI: 10.1016/j.str.2022.02.005
• 发表时间: 2022-05-05
• 期刊: STRUCTURE
• 影响因子: 5.7
• 作者: Kantsadi, Anastassia L.;Hatzopoulos, Georgios N.;Gonczy, Pierre;Vakonakis, Ioannis
• 通讯作者: Vakonakis, Ioannis