Appel L F, Prout M, Abu-Shumays R, Hammonds A, Garbe J C, Fristrom D, Fristrom J
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):4937-41. doi: 10.1073/pnas.90.11.4937.
The Stubble-stubbloid (Sb-sbd) gene is required for hormone-dependent epithelial morphogenesis of imaginal discs of Drosophila, including the formation of bristles, legs, and wings. The gene has been cloned by using Sb-sbd-associated DNA lesions in a 20-kilobase (kb) region of a 263-kb genomic walk. The region specifies an approximately 3.8-kb transcript that is induced by the steroid hormone 20-hydroxyecdysone in imaginal discs cultured in vitro. The conceptually translated protein is an apparent 786-residue type II transmembrane protein (N terminus in, C terminus out), including an intracellular N-terminal domain of at least 35 residues and an extracellular C-terminal trypsin-like serine protease domain of 244 residues. Sequence analyses indicate that the Sb-sbd-encoded protease could activate itself by proteolytic cleavage. Consistent with the cell-autonomous nature of the Sb-sbd bristle phenotype, a disulfide bond between cysteine residues in the noncatalytic N-terminal fragment and the C-terminal catalytic fragment could tether the protease to the membrane after activation. Both dominant Sb and recessive sbd mutations affect the organization of microfilament bundles during bristle morphogenesis. We propose that the Sb-sbd product has a dual function. (i) It acts through its proteolytic extracellular domain to detach imaginal disc cells from extracellular matrices, and (ii) it transmits an outside-to-inside signal to its intracellular domain to modify the cytoskeleton and facilitate cell shape changes underlying morphogenesis.
须茬-类须茬(Sb-sbd)基因是果蝇成虫盘激素依赖性上皮形态发生所必需的,包括刚毛、腿和翅膀的形成。通过在263千碱基(kb)基因组步移的20千碱基(kb)区域中使用与Sb-sbd相关的DNA损伤克隆了该基因。该区域指定了一个约3.8千碱基的转录本,在体外培养的成虫盘中由类固醇激素20-羟基蜕皮激素诱导产生。从概念上翻译的蛋白质是一种明显的786个残基的II型跨膜蛋白(N端在内,C端在外),包括至少35个残基的细胞内N端结构域和244个残基的细胞外C端胰蛋白酶样丝氨酸蛋白酶结构域。序列分析表明,Sb-sbd编码的蛋白酶可以通过蛋白水解切割来激活自身。与Sb-sbd刚毛表型的细胞自主性一致,非催化N端片段和C端催化片段中的半胱氨酸残基之间的二硫键可以在激活后将蛋白酶 tether 到膜上。显性Sb和隐性sbd突变都会影响刚毛形态发生过程中微丝束的组织。我们提出,Sb-sbd产物具有双重功能。(i)它通过其蛋白水解细胞外结构域作用,使成虫盘细胞与细胞外基质分离,(ii)它将细胞外信号传递到其细胞内结构域,以修饰细胞骨架并促进形态发生过程中潜在的细胞形状变化。 (注:“tether”这个词在原文语境中可能是一个专业术语,这里直接保留英文未翻译,需结合具体医学知识进一步确定其准确含义以便完整准确翻译)