Ruiz María-Fernanda, Sánchez Lucas
Centro de Investigaciones Biologicas (CSIC), Madrid, Spain.
Int J Dev Biol. 2010;54(4):627-33. doi: 10.1387/ijdb.092917fr.
The gene transformer (tra) is the key regulatory memory device for sex determination in tephritid insects. The present manuscript addressed the question about the functional conservation of the tephritid Anastrepha Transformer protein to direct somatic sexual development in Drosophila (Drosophilidae). The transformer cDNA of Anastrepha encoding the putative full-length Tra protein was cloned in pUAST and introduced into Drosophila melanogaster. To express this protein, the GAL4-UAS system was used. The Anastrepha Tra protein induced the female-specific splicing of both dsx and fru pre-mRNAs in Drosophila XY male flies, so that these became transformed into females, though this transformation was incomplete (the sexually dimorphic foreleg basitarsus and the external terminalia were monitored). It was found that the degree of female transformation directly depended on the dose of Anastrepha tra and Drosophila transformer-2 (tra-2) genes, and that the Anastrepha Tra-Drosophila Tra2 complex is not as efficient as the Drosophila Tra-Tra2 complex at inducing the female-specific splicing of Drosophila dsx pre-mRNA. This can explain why the Anastrepha Tra protein cannot fully substitute for the endogenous Drosophila Tra protein.
基因transformer(tra)是实蝇科昆虫性别决定的关键调控记忆元件。本论文探讨了实蝇科果实蝇属的transformer蛋白在指导果蝇(果蝇科)体细胞性发育方面功能保守性的问题。将编码推定全长Tra蛋白的果实蝇属transformer cDNA克隆到pUAST载体中,并导入黑腹果蝇。为了表达该蛋白,使用了GAL4-UAS系统。果实蝇属Tra蛋白诱导了果蝇XY雄蝇中dsx和fru前体mRNA的雌性特异性剪接,从而使这些雄蝇转变为雌蝇,不过这种转变并不完全(监测了两性异形的前足基跗节和外部生殖器)。研究发现,雌性转变的程度直接取决于果实蝇属tra和果蝇transformer-2(tra-2)基因的剂量,并且果实蝇属Tra-果蝇Tra2复合物在诱导果蝇dsx前体mRNA的雌性特异性剪接方面不如果蝇Tra-Tra2复合物有效。这可以解释为什么果实蝇属Tra蛋白不能完全替代内源性果蝇Tra蛋白。