[
International Worm Meeting,
2005]
The smooth sinusoidal movements of C. elegans are effected by 95 obliquely striated bodywall muscle cells. There are also non-striated single-sarcomere muscles; the pharyngeal, intestinal, anal and sex-specific muscles. Force generated by the myofilament lattice of bodywall muscle is transmitted to the external cuticle through the attachment structures of the dense bodies, M-lines and fibrous organelles. Attachment structures are also present in the single-sarcomere muscle cells. Additional components of the lattice and associated attachment structures may be identified by investigating the subcellular localisation of endogenous proteins fused to GFP. The twin resources of the Promoterome and ORFeome are being used to construct Promoter::ORF::GFP fusions in a high-throughput manner using Multisite Gateway recombinational cloning. Transgenic lines are generated using these constructs to study the expression patterns of ORF::GFP fusion proteins in vivo. The genes selected for inclusion in this project were chosen because their respective promoters had been shown previously to drive expression of reporter genes in muscle cells. Including ORFs within Promoter::ORF::GFP constructs may reveal the subcellular localisation of the native protein. The genes W05F2.4, F07C3.4 and F02A9.3 were used to establish procedures and expression patterns will be presented. Relative localisation of the fusion proteins with well-known components of muscle structures will be investigated using reporter genes of differing colours. This would also facilitate visualisation of the various components of the contractile apparatus and associated attachment structures assembling throughout development. After initially identifying novel components, techniques such as RNAi and yeast two-hybrid experiments may be employed to clarify the roles and interactions of these proteins.