Burnette J M, Hatton A R, Lopez A J
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennyslvania 15213, USA.
Genetics. 1999 Apr;151(4):1517-29. doi: 10.1093/genetics/151.4.1517.
Alternatively spliced Ultrabithorax mRNAs differ by the presence of internal exons mI and mII. Two approaches were used to identify trans-acting factors required for inclusion of these cassette exons. First, mutations in a set of genes implicated in the control of other alternative splicing decisions were tested for dominant effects on the Ubx alternative splicing pattern. To identify additional genes involved in regulation of Ubx splicing, a large collection of deficiencies was tested first for dominant enhancement of the haploinsufficient Ubx haltere phenotype and second for effects on the splicing pattern. Inclusion of the cassette exons in Ubx mRNAs was reduced strongly in heterozygotes for hypomorphic alleles of hrp48, which encodes a member of the hnRNP A/B family and is implicated in control of P-element splicing. Significant reductions of mI and mII inclusion were also observed in heterozygotes for loss-of-function alleles of virilizer, fl(2)d, and crooked neck. The products of virilizer and fl(2)d are also required for Sxl autoregulation at the level of splicing; crooked neck encodes a protein with structural similarities to yeast-splicing factors Prp39p and Prp42p. Deletion of at least five other loci caused significant reductions in the inclusion of mI and/or mII. Possible roles of identified factors are discussed in the context of the resplicing strategy for generation of alternative Ubx mRNAs.
选择性剪接的超双胸复合体(Ultrabithorax)mRNA因内部外显子mI和mII的存在与否而有所不同。我们采用了两种方法来鉴定包含这些盒式外显子所需的反式作用因子。首先,对一组与其他可变剪接决定控制相关的基因突变进行测试,以检测其对超双胸复合体可变剪接模式的显性作用。为了鉴定参与超双胸复合体剪接调控的其他基因,首先对大量缺失突变体进行测试,以检测其对单倍剂量不足的超双胸复合体平衡棒表型的显性增强作用,其次检测其对剪接模式的影响。对于编码hnRNP A/B家族成员且与P因子剪接控制有关的hrp48亚效等位基因的杂合子,超双胸复合体mRNA中盒式外显子的包含率大幅降低。在雄性化因子(virilizer)、fl(2)d和弯颈(crooked neck)功能缺失等位基因的杂合子中,也观察到mI和mII包含率的显著降低。雄性化因子和fl(2)d的产物在剪接水平上对性致死基因(Sex-lethal, Sxl)的自我调节也是必需的;弯颈编码一种与酵母剪接因子Prp39p和Prp42p结构相似的蛋白质。至少五个其他位点的缺失导致mI和/或mII的包含率显著降低。我们在产生可变超双胸复合体mRNA的重新剪接策略的背景下讨论了已鉴定因子的可能作用。