Three-dimensional reconstruction of abscission

脱离的三维重建

• 批准号: 198846519
• 负责人: Professor Dr. Thomas Müller-Reichert
• 金额: --
• 依托单位: Experimentelles Zentrum
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/198846519?language=en
• 关键词: Three; dimensional; reconstruction; abscission

Abscission is the final stage of cytokinesis, when the narrow intercellular bridge connecting the two post-mitotic daughter cells is severed. Recently, we discovered helices of ESCRT-III-dependent fila-ments forming within the intercellular bridges of HeLa cells that could drive cortical constriction during abscission. The first goal of this research proposal is to study the relative contributions of CEP55 and ALIX, two potential upstream targeting components of the ESCRT-III complex, and Spastin, to abscission in HeLa cells using electron tomography. The second goal is to extend our analysis of abscission to the early Caenorhabditis elegans embryo. We want to answer the basic question, is our suggested model of abscission conserved in other systems? Using the worm, we propose to generate reference 3D models of intermediate stages of abscission in wild-type embryos, characterize the role of C. elegans ESCRT-III components by RNA-mediated interference (RNAi), and perform electron tomographic analyses of staged mutant embryos. It is anticipated that this comparative approach will lead to further fundamental structural and mechanistic insights into the process of abscission.

分离是细胞质分裂的最后阶段，连接两个有丝分裂后子细胞的狭窄细胞间桥被切断。最近，我们发现HeLa细胞的细胞间桥内形成escrt - iii依赖性丝的螺旋，可以在脱落过程中驱动皮质收缩。本研究计划的第一个目标是利用电子断层扫描研究CEP55和ALIX （ESCRT-III复合体的两个潜在上游靶向成分）和Spastin对HeLa细胞脱落的相对贡献。第二个目标是将我们对分离的分析扩展到秀丽隐杆线虫的早期胚胎。我们想要回答一个基本的问题，我们建议的分离模型在其他系统中是否守恒？利用线虫，我们建议在野生型胚胎中生成分离中间阶段的参考3D模型，通过rna介导干扰（RNAi）表征秀丽隐杆线虫ESCRT-III成分的作用，并对分阶段突变胚胎进行电子断层扫描分析。预计这种比较方法将导致对脱落过程进一步的基本结构和机械见解。