P element insertion-dependent gene activation in the Drosophila eye
P element insertion-dependent gene activation in the Drosophila eye
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DOI：
10.1073/pnas.94.10.5195
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发表时间：
1997-05-13
影响因子：
11.1
通讯作者：
Rubin, GM
中科院分区：
文献类型：
--
作者：
Hay, BA;Maile, R;Rubin, GM
了解基因的功能可以通过多种方式获得，包括功能丧失表型、序列相似性、表达模式以及其错误表达的后果。当其他方法无法解释时，对基因在异常时间、地点或水平表达所产生的表型进行分析，可以为基因的功能提供线索。在这里，我们报告，眼睛特异性，增强子启动子存在于P元件表达载体pGMR能够驱动高水平的表达在眼睛的基因附近的P元件插入的网站。细胞命运决定、分化、增殖和死亡对于正常的眼睛发育是必不可少的。因此，考虑到眼睛对于成人的生存力和生育力的可分配性，进行基因组中基因的显著部分的眼睛特异性错误表达的能力应该为鉴定这些过程的调节剂提供有力的方法。为了验证这一想法，我们对调节细胞死亡的基因进行了两次过表达筛选。我们在野生型背景中筛选了插入依赖的显性表型，并筛选了收割机过表达诱导的小眼表型的显性修饰剂。鉴定了多个染色体基因座，包括在5'端插入HID（一种有效的细胞凋亡诱导剂）和在5'端插入DIAP 1（一种细胞死亡抑制剂）。为了便于克隆P元件插入附近的基因，创建了新的错误表达载体。一个屏幕与这些载体之一确定鹰作为一个抑制剂的粗糙的眼睛表型与激活的Ras 1基因的过表达。
Insights into the function of a gene can be gained in multiple ways, including loss-of-function phenotype, sequence similarity, expression pattern, and by the consequences of its misexpression. Analysis of the phenotypes produced by expression of a gene at an abnormal time, place, or level may provide clues to a gene's function when other approaches are not illuminating. Here we report that an eye-specific, enhancer-promoter present in the P element expression vector pGMR is able to drive high level expression in the eye of genes near the site of P element insertion. Cell fate determination, differentiation, proliferation, and death are essential for normal eye development. Thus the ability to carry out eye-specific misexpression of a significant fraction of genes in the genome, given the dispensability of the eye for viability and fertility of the adult, should provide a powerful approach for identifying regulators of these processes. To test this idea we carried out two overexpression screens for genes that function to regulate cell death. We screened for insertion-dependent dominant phenotypes in a wild-type background, and for dominant modifiers of a reaper overexpression-induced small eye phenotype. Multiple chromosomal loci were identified, including an insertion 5' to hid, a potent inducer of apoptosis, and insertions 5' to DIAP1, a cell death suppressor. To facilitate the cloning of genes near the P element insertion new misexpression vectors were created. A screen with one of these vectors identified eagle as a suppressor of a rough eye phenotype associated with overexpression of an activated Ras1 gene.