Drosophila singed, a fascin homolog, is required for actin bundle formation during oogenesis and bristle extension.
Drosophila singed mutants were named for their gnarled bristle phenotype but severe alleles are also female sterile. Recently, singed protein was shown to have 35% peptide identity with echinoderm fascin. Fascin is found in actin filament bundles in microvilli of sea urchin eggs and in filopodial extensions in coelomocytes. We show that Drosophila singed is required for actin filament bundle formation in the cytoplasm of nurse cells during oogenesis; in severe mutants, the absence of cytoplasmic actin filament bundles allows nurse cell nuclei to lodge in ring canals and block nurse cell cytoplasm transport. Singed is also required for organized actin filament bundle formation in the cellular extension that forms a bristle; in severe mutants, the small disorganized actin filament bundles lack structural integrity and allow bristles to bend and branch during extension. Singed protein is also expressed in migratory cells of the developing egg chamber and in the socket cell of the developing bristle, but no defect is observed in these cells in singed mutants. Purified, bacterially expressed singed protein bundles actin filaments in vitro with the same stoichiometry reported for purified sea urchin fascin. Singed-saturated actin bundles have a molar ratio of singed/actin of approximately 1:4.3 and a transverse cross-banding pattern of 12 nm seen using electron microscopy. Our results suggest that singed protein is required for actin filament bundle formation and is a Drosophila homolog of echinoderm fascin.
Cant
K
K
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
Knowles
B A
BA
Mooseker
M S
MS
Cooley
L
L
D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism*; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D004330:Drosophila / Q000196:embryology / Q000378:metabolism*; D005260:Female; D007301:Insect Hormones / Q000276:immunology / Q000378:metabolism*; D008840:Microfilament Proteins / Q000378:metabolism*; D009866:Oogenesis; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D011994:Recombinant Proteins / Q000378:metabolism; D017385:Sequence Homology
D000199:Actins; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D007301:Insect Hormones; D008840:Microfilament Proteins; D011994:Recombinant Proteins; C020324:fascin