Determination of the mobility of novel and established Caenorhabditis elegans sarcomeric proteins in vivo.
A screen was instigated to identify novel protein components of the Caenorhabditis elegans sarcomere. The subcellular localisation of full-length GFP fusion proteins was examined, in transgenic animals, for 62 essentially uncharacterized genes thought to be expressed within bodywall muscle cells. Three genes, T03G6.3, C46G7.2 and K04A8.6, were identified for further study. K04A8.6::GFP only displayed a regular sarcomeric distribution sporadically. However, C46G7.2::GFP localised to the centre of A-bands and dense bodies and T03G6.3::GFP localised in the I-band, of the bodywall muscle sarcomeres, consistently. This success with such a small screen suggests that there are further minor components of the C. elegans sarcomere yet to be discovered. Fluorescence Recovery After Photobleaching (FRAP) was applied to live transgenic individuals to assess the mobility of T03G6.3 and C46G7.2 and other well-known constituents of the sarcomere in vivo. Proteins associated with the thin filaments showed dynamic exchange whilst those associated with thick filaments appeared more static. This is the first demonstration that there are sarcomeric proteins in C. elegans muscle cells in dynamic exchange and that the rates of exchange in vivo correspond in general terms with observations in other experimental systems.
Institute of Integrative and Comparative Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK.
Shipa R
SR
Ghosh
Ian A
IA
Hope
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D017238:Genes, Helminth; D049452:Green Fluorescent Proteins; D018613:Microscopy, Confocal; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012518:Sarcomeres / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure
C418503:C46G7.2 protein, C elegans; D029742:Caenorhabditis elegans Proteins; D009124:Muscle Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins