Functional analysis of the single calmodulin gene in the nematode Caenorhabditis elegans by RNA interference and 4-D microscopy.
Calmodulin (CaM), a small calcium-binding protein, is the key mediator of numerous calcium-induced changes in cellular activity. Its ligands include enzymes, cytoskeletal proteins and ion channels, identified in large part by biochemical and cell biological approaches. Thus far it has been difficult to assess the function of CaM genetically, because of the maternal supply in Drosophila and the presence of at least three nonallelic genes in vertebrates. Here we use the unique possibility offered by the C. elegans model system to inactivate the single CaM gene (cmd-1) through RNA interference (RNAi). We show that the RNAi microinjection approach results in a severe embryonic lethal phenotype. Embryos show disturbed morphogenesis, aberrant cell migration patterns, a striking hyperproliferation of cells and multiple defects in apoptosis. Finally, we show that RNAi delivery by the feeding protocol does not allow the efficient silencing of the CaM gene obtained by microinjection. General differences between the two delivery methods are discussed.
Karabinos
Anton
A
Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Goettingen, Germany.
Büssing
Ingo
I
Schulze
Ekkehard
E
Wang
Jian
J
Weber
Klaus
K
Schnabel
Ralf
R
D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000033:anatomy & histology* / Q000196:embryology / Q000235:genetics; D002147:Calmodulin / Q000235:genetics*; D002455:Cell Division / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology; D018507:Gene Expression Regulation, Developmental; D008845:Microinjections; D008853:Microscopy; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000502:physiology; D010641:Phenotype; D034622:RNA Interference; D020539:Sequence Analysis, Protein
D002147:Calmodulin