Kraus K W, Lee Y H, Lis J T, Wolfner M F
Section of Genetics and Development, Cornell University, Ithaca, New York 14853.
Mol Cell Biol. 1988 Nov;8(11):4756-64. doi: 10.1128/mcb.8.11.4756-4764.1988.
The sex of Drosophila melanogaster is determined by a hierarchy of genes. The ultimate targets of this regulatory hierarchy are the genes encoding terminal differentiation products of one sex. For one of the best-characterized target genes, that encoding female-specific yolk protein 1 (YP1), sex-specific transcriptional controls have been clearly demonstrated. In addition, sex-specific posttranscriptional controls were suggested from experiments in which YP1 RNA was induced in males with hormones. To determine whether males can efficiently process and translate a transcript which is normally found only in females, we used a non-sex-specific promoter, the hsp70 gene promoter, to drive YP1 gene transcription in germ line transformed males. The efficiency of expression of the YP1 gene at levels of RNA splicing, translation, and protein secretion in these males was compared with that in wild-type females. These experiments show that there are no sex-specific posttranscriptional controls operating to limit the production of secreted YP1 in males. Promoters containing different numbers of heat shock elements were tested for their ability to drive YP1 gene transcription in males. These results show that incompatibility between the hsp70 gene heat shock elements and the YP1 gene promoter can be overcome by increasing the amount of hsp70 gene sequence up or downstream of the TATA box. In the course of this study, two vectors useful for placing genes under heat shock regulation were constructed. One of these vectors is designed so that the heat-induced transcript produced is the "authentic" primary transcript; it should be useful for studies of posttranscriptional regulation.
黑腹果蝇的性别由一系列基因决定。这一调控层级的最终靶点是编码某一性别的终末分化产物的基因。对于其中一个特征最为明确的靶点基因,即编码雌性特异性卵黄蛋白1(YP1)的基因,已经明确证实了性别特异性转录调控。此外,在利用激素诱导雄性产生YP1 RNA的实验中,也提示了性别特异性转录后调控。为了确定雄性是否能够有效加工和翻译通常仅在雌性中发现的转录本,我们使用了一个非性别特异性启动子,即热休克蛋白70(hsp70)基因启动子,来驱动种系转化雄性中的YP1基因转录。将这些雄性中YP1基因在RNA剪接、翻译和蛋白质分泌水平上的表达效率与野生型雌性中的进行了比较。这些实验表明,不存在性别特异性转录后调控来限制雄性中分泌型YP1的产生。测试了含有不同数量热休克元件的启动子驱动雄性中YP1基因转录的能力。这些结果表明,通过增加TATA框上游或下游的hsp70基因序列的量,可以克服hsp70基因热休克元件与YP1基因启动子之间的不相容性。在这项研究过程中,构建了两个可用于将基因置于热休克调控之下的载体。其中一个载体的设计使得热诱导产生的转录本是“真实的”初级转录本;它应该对转录后调控的研究有用。