The role of the centriolar CEP350 network in centrobin removal from centrioles and distal appendage/CEP19 recruitment

中心粒CEP350网络在中心粒从中心粒和远端附属物去除中心蛋白中的作用/CEP19招募

• 批准号: 421323611
• 负责人: Professorin Dr. Gislene Pereira
• 金额: --
• 依托单位: Zentrum für Molekulare Biologie (ZMBH)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/421323611?language=en
• 关键词: role; centriolar; CEP350; network; centrobin

The centrosome is vital for cellular organization and division, and defects in its structure and function are linked to various human diseases. This proposal focuses on the centrosome protein CEP350 that is critical for controlling centriole length, stability, assembling subdistal (SDA) and distal (DA) appendages and centriole recruitment of CEP19. DA assembly and CEP19 recruitment are essential for ciliogenesis, and defects in their assembly are linked to various diseases such as ciliopathies and morbid obesity (CEP19). CEP350 also regulates the centriolar protein centrobin, which is necessary for proper centrosome function and is specifically removed from maturing centrioles by proteolysis. Dysregulation of centrobin removal is linked to Mulibrey nanism and cancer. Despite the importance of CEP350 for DA assembly, CEP19 recruitment and centrobin removal, its molecular functions are poorly understood. Understanding how CEP350 regulates these processes is critical for elucidating key open questions in centrosome biology and disease mechanisms and therefore stands in the focus of this proposal. Based on our preliminary findings, we propose that CEP350 is a multifunctional protein that cooperates with DISCO network components (several centriolar proteins needed together with CEP350 for DA assembly), DA proteins and CEP19 in order to achieve its functions. Our study, which combines the expertise of the laboratories of Profs. Pereira and Schiebel in centriole biology, CEP350 analysis, DA assembly and ciliogenesis, focuses on two major goals. Firstly, we aim to explore the detailed mechanisms of centrobin removal from centrioles by CEP350, SDAs, the E3 ligase TRIM37 and the by us newly identified kinase CRM5. In addition, we will also investigate whether centrobin removal is essential for DA assembly. Secondly, we will investigate how CEP350 interacting proteins and mother centriole specific modifications regulate DA assembly and CEP19 centriole recruitment at maturing centrioles. To achieve these goals, we will use various techniques, including gene knockouts, signal intensity measurements, super-resolution/expansion microscopy, electron microscopy, Minflux nanoscopy and mass spectrometry analysis of phosphorylation sites in a subset of CEP350 interacting proteins. By uncovering how CEP350 controls centrobin and DA assembly pathways, our study will help answering critical questions in centrosome biology, cilia assembly and how defects lead to disease.

中心体对细胞组织和分裂至关重要，其结构和功能的缺陷与各种人类疾病有关。该提案的重点是中心体蛋白CEP 350，该蛋白对于控制中心粒长度、稳定性、近远端（SDA）和远端（DA）附件的组装以及CEP 19的中心粒募集至关重要。DA组装和CEP 19募集对于纤毛发生是必不可少的，并且其组装中的缺陷与各种疾病如纤毛病和病态肥胖（CEP 19）相关。CEP 350还调节中心粒蛋白质centrobin，其对于适当的中心体功能是必需的，并且通过蛋白水解从成熟的中心粒中特异性地去除。centrobin去除的失调与Mulibrey nanism和癌症有关。尽管CEP 350对于DA组装、CEP 19募集和centrobin去除的重要性，但其分子功能知之甚少。了解CEP 350如何调节这些过程对于阐明中心体生物学和疾病机制中的关键开放问题至关重要，因此是本提案的重点。基于我们的初步研究结果，我们提出，CEP 350是一个多功能的蛋白质，合作DISCO网络组件（需要与CEP 350一起DA组装的几个中心粒蛋白），DA蛋白和CEP 19，以实现其功能。我们的研究，结合了教授实验室的专业知识。佩雷拉和Schiebel在中心粒生物学、CEP 350分析、DA组装和纤毛发生中，专注于两个主要目标。首先，我们的目的是探讨CEP 350，SDAs，E3连接酶TRIM 37和我们新发现的激酶CRM 5从中心粒去除centrobin的详细机制。此外，我们还将调查是否centrobin去除是必不可少的DA大会。其次，我们将研究如何CEP 350相互作用的蛋白质和母亲的中心粒特异性修饰调节DA组装和CEP 19中心粒招募成熟的中心粒。为了实现这些目标，我们将使用各种技术，包括基因敲除，信号强度测量，超分辨率/放大显微镜，电子显微镜，Minflux nanoscopy和质谱分析的磷酸化位点在一个子集的CEP 350相互作用蛋白。通过揭示CEP 350如何控制centrobin和DA组装途径，我们的研究将有助于回答中心体生物学，纤毛组装以及缺陷如何导致疾病的关键问题。