Cant K, Knowles B A, Mooseker M S, Cooley L
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
J Cell Biol. 1994 Apr;125(2):369-80. doi: 10.1083/jcb.125.2.369.
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.
果蝇“烧焦”突变体因其刚毛扭曲的表型而得名,但严重的等位基因也会导致雌性不育。最近研究表明,“烧焦”蛋白与棘皮动物的肌动蛋白束蛋白有35%的肽段一致性。肌动蛋白束蛋白存在于海胆卵微绒毛中的肌动蛋白丝束以及体腔细胞的丝状伪足延伸中。我们发现,果蝇“烧焦”基因对于卵子发生过程中滋养细胞胞质内肌动蛋白丝束的形成是必需的;在严重突变体中,胞质内缺乏肌动蛋白丝束会使滋养细胞核嵌入环管并阻碍滋养细胞胞质的运输。“烧焦”基因对于形成刚毛的细胞延伸中有序的肌动蛋白丝束形成也是必需的;在严重突变体中,小而无序的肌动蛋白丝束缺乏结构完整性,使得刚毛在延伸过程中弯曲和分支。“烧焦”蛋白也在发育中的卵室的迁移细胞以及发育中的刚毛的窝细胞中表达,但在“烧焦”突变体的这些细胞中未观察到缺陷。纯化的、细菌表达的“烧焦”蛋白在体外能使肌动蛋白丝成束,其化学计量与纯化的海胆肌动蛋白束蛋白相同。饱和“烧焦”的肌动蛋白束中“烧焦”蛋白与肌动蛋白的摩尔比约为1:4.3,用电子显微镜观察呈现12纳米的横向交叉带纹模式。我们的结果表明,“烧焦”蛋白是肌动蛋白丝束形成所必需的,并且是棘皮动物肌动蛋白束蛋白在果蝇中的同源物。