Ability of scalloped deletion constructs to rescue sd mutant wing phenotypes in Drosophila melanogaster
Ability of scalloped deletion constructs to rescue sd mutant wing phenotypes in Drosophila melanogaster
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作者：
Chow, L;Berube, J;Bell, JB
扇贝(SD)和残留(VG)蛋白在物理上相互作用，形成一个选择器复合体，激活参与黑腹果蝇翅膀发育的基因。SD属于一个保守的转录因子家族，含有TEA/ATTS DNA结合基序。VG也是一种核蛋白，为SD VG复合体提供激活功能。茶树DNA结合区和SD的VG相互作用域(VID)已被鉴定和描述。然而，它们，可能还有SD的其他功能域，还没有在体内得到彻底的表征。在这里，编码不同截短的SD的转基因构建物被用来评估它们各自拯救两个可存活的SD隐性突变(SD(ETX4)和SD(58d))突变翼型的能力。对产生的转基因菌株也进行了诱导SD进一步表达的能力测试，这是全长SD所具有的能力。SD的功能解剖证实了特定的区域对于翅膀的发育是必要的，并提供了关于SD VG复合体如何发挥作用以促进翅膀命运的进一步信息。先前的实验表明，全长SD的表达可以导致显性的负翼表型。我们表明，表达删除SDDNA结合域的构建体也可以在两个测试者SD菌株的背景中导致显性阴性表型。相反，删除VID的SD构建体在两个测试器背景中都不会对翅膀表型产生影响。最后，对于正常的翅膀发育来说，N端的很大一部分SD似乎是可有可无的，因为在我们的分析中，这种结构的行为与全长SD相同。
Scalloped (SD) and Vestigial (VG) proteins physically interact to form a selector complex that activates genes involved in wing development in Drosophila melanogaster. SD belongs to a conserved family of transcription factors containing the TEA/ATTS DNA-binding motif. VG is also a nuclear protein providing the activator function for the SD VG complex. The TEA DNA-binding domain and the VG interacting domain (VID) of SD have been previously identified and described. However, they, and possibly other functional domains of SD, have not been thoroughly characterized in vivo. Herein, transgenic constructs encoding various truncations of SD were used to assess their respective ability to rescue the mutant wing phenotype of two viable sd recessive mutations (sd(ETX4) and sd(58d)). The transgenic strains produced were also tested for the ability to induce further sd expression, an ability possessed by full length SD. The functional dissection of SD confirms that specific regions are necessary for wing development and provides further information as to how the SD VG complex functions to promote wing fate. Previous experiments have shown that expression of full length SD can cause a dominant negative wing phenotype. We show that expression of constructs that delete the SD DNA-binding domain can also cause a dominant negative phenotype in a background with either of the two tester sd strains. In contrast, SD constructs that delete the VID have no effect on the wing phenotype in either tester background. Finally, a significant portion of SD at the N-terminal end appears to be dispensable with respect to normal wing development, as this construct behaves the same as full length SD in our assays.