Analysis of in vivo consequences of disease-relvant point mutations in p97 (VCP) in Dictostelium discoideum, a simple and powerful model system for functional genomics

盘网柄菌 p97 (VCP) 疾病相关点突变的体内后果分析，盘柄菌是一种简单而强大的功能基因组模型系统

• 批准号: 149382680
• 负责人: Professor Dr. Ludwig Eichinger
• 金额: --
• 依托单位: Institut für Luft- und Raumfahrtmedizin
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/149382680?language=en
• 关键词: Analysis; vivo; consequences; disease; relvant

p97 (VCP or valosin-containing protein in mammals and Cdc48p in yeast) is a ubiquitously expressed and evolutionarily highly conserved member of the magnesium-dependent Walker P-loop AAA-ATPases. Mutations in the human p97 gene on chromosome 9p13-p12 cause the autosomal dominant inclusion body myopathy associated with Paget disease of the bone and frontotemporal dementia or IBMPFD. In this project we will study the function of p97 in D. discoideum, a simple and powerful haploid model system for functional genomics as well as an established model for autophagic processes. We have already successfully generated Dictyostelium strains that express endogenous p97 as well as wildtype or mutant R155C p97 fused to GFP or RFP. The latter strain mimics the genetic situation in human IBMPFD patients. Special emphasis of our analyses will be on protein aggregate formation, proteasomal and autophagic dysfunction. We will search for novel p97 binding partners by carrying out immunoprecipitations followed by mass spectrometry. To identify diseaserelevant intracellular signalling pathways, comparative microarray analyses in conjunction with sophisticated bioinformatic methods will be performed. The main goal of this project will be the elucidation of the molecular and cellular consequences of the R155C mutation in Proposal for a DFG-Research Unit: Myofibrillar Myopathies 68 p97 in the Dictyostelium model system and comparison with the human and murine IBMPFD pathology.

p97（哺乳动物中的VCP或含缬氨酸蛋白，酵母中的Cdc48p）是镁依赖性Walker P-loop aaa - atp酶中普遍表达且进化高度保守的成员。染色体9p13-p12上的人类p97基因突变导致常染色体显性包涵体肌病与骨Paget病和额颞叶痴呆或IBMPFD相关。在本项目中，我们将研究p97在D. disideum中的功能，D. disideum是一个简单而强大的功能基因组学单倍体模型系统，也是一个已建立的自噬过程模型。我们已经成功地产生了Dictyostelium菌株，表达内源性p97以及野生型或突变型R155C p97融合到GFP或RFP。后一种菌株模拟了人类IBMPFD患者的遗传情况。我们的分析将特别强调蛋白质聚集形成，蛋白酶体和自噬功能障碍。我们将通过进行免疫沉淀和质谱分析来寻找新的p97结合伙伴。为了确定与疾病相关的细胞内信号通路，将结合复杂的生物信息学方法进行比较微阵列分析。本项目的主要目标是阐明dfg研究单元提案中R155C突变的分子和细胞后果：肌纤原性肌病68 p97在Dictyostelium模型系统中，并与人类和小鼠IBMPFD病理进行比较。