Characterization of LEM-domain Proteins in Drosophila melanogaster and their relevance regarding human laminopathic diseases

• Applicant: Dr. Nicole Wagner
• Subject Area: Cell Biology
• Term: from 2006 to 2010
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 29303608

The nuclear envelope is composed of three distinct membrane domains, the inner and outer nuclear membrane and the wall of nuclear pore complexes. A characteristic feature for the inner nuclear membrane is the nuclear lamina and a subset of inner nuclear membrane proteins, including LEM-domain proteins. A growing number of human inherited diseases known as laminopathies, are caused by mutations in genes encoding lamin A/C and LEM-domain proteins. The complexity of LEM-domain protein composition in Drosophila is comparable to that in human, therefore the fruitfly present a powerful genetic system for analyzing the functional relationships among LEM-domain proteins, lamins and BAF, as well as the mechanisms underlying laminopathic diseases. A combination of cell biological, genetic and biochemical approaches will be used to address fundamental questions concerning the structure, organization, interactions and basic functions of LEM-domain proteins in Drosophila. The developmental and possible cell type specific expression pattern of LEM-domain proteins will be analyzed using proteinblot- and whole mount in situ hybridization techniques. To determine the interaction of the Drosophila LEM-domain proteins with lamins and BAF, co-immunoprecipitations and blot overlay experiments will be performed. To identify potential novel binding partners, the Yeast Two-Hybrid System will be used. Potential interactions will be verified using blot overlay experiments and RNAi experiments. To analyze the (possible overlapping) functions of the Drosophila LEM-domain proteins, these proteins as well as their potential binding partners will be simultaneously down regulated in different combinations in Drosophila cells and during Drosophila development by RNAi.

• DFG Programme: Research Fellowships
• International Connection: Sweden
• Hosts: Professor Dr. Georg Krohne; Professor Dr. Christos Samakovlis