Analysis of new muscle specific genes/proteins and their function in sarcomere assembly in C. elegans using SAGE and Affymetrix Microarray Chip technology including Bioinformatics

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The aim of my research proposal is to identify novel genes/proteins involved in sarcomere assembly in C. elegans. With the fellowship I am applying for, I will work on a project in the laboratory of Prof. Don Moerman at the University of British Columbia, Vancouver. His lab focuses on muscle development and specifically on the problem of muscle sarcomere assembly in C. elegans. This system is an excellent in vivo model to study the assembly of integrin-containing adhesion structures. Recently, a large number of muscle specific genes have been identified by serial analysis of gene expression (SAGE) using C. elegans muscle specific RNA at the Michael Smith Genome Sciences Center, Vancouver. I will use this database to help find new genes/proteins placed in adhesion complexes and to determine their function during sarcomere assembly. The specific goals of the proposed research are as follows: 1) Determination of the expression pattern of different candidate genes in C. elegans focusing on subcellular localization. 2) Generating C. elegans mutant animals for the genes of interest using RNAi or a mutagenesis screen. 3) Phenotyp analysis of mutant animals focusing on sarcomere development. 4) Biochemical analysis of interesting target proteins. Since there are many genes having human homologues with unknown function the findings will be very useful as myofilament lattice assembly in the nematode is remarkably similar to the assembly of focal adhesions in mammalian cell culture.