Lossky M, Wensink P C
Rosenstiel Center, Brandeis University, Waltham, Massachusetts 02254-9110, USA.
Mol Cell Biol. 1995 Dec;15(12):6943-52. doi: 10.1128/MCB.15.12.6943.
The divergently transcribed yolk protein genes (Yp1 and Yp2) of Drosophila melanogaster are expressed only in adult females, in fat body tissue and in ovarian follicle cells. Using an in vitro transcription assay, we have identified a single 12-bp DNA element that activates transcription from the promoters of both Yp genes. In vivo, this regulatory element is tissue specific: it activates transcription of Yp1 and Yp2 reporter genes in follicle cells but has no detectable effect in fat body or other tissues. The sequence of the element consists of two recognition sites for the GATA family of transcription factors. We show that among the Drosophila genes known to encode GATA factors, only dGATAb is expressed in ovaries. The single transcript that we detect in ovaries is alternatively spliced or initiated to produce an ovary-specific isoform of the protein. Bacterially expressed dGATAb binds to the 12-bp element; a similar binding activity is also present in the Kc0 nuclear extracts used for in vitro transcription assays. These in vitro and in vivo results lead us to propose that dGATAb makes several developmentally regulated products, one of which is a follicle cell-specific protein activating transcription of Yp1 and Yp2 from a known regulatory element.
果蝇的卵黄蛋白基因(Yp1和Yp2)呈发散转录,仅在成年雌性果蝇的脂肪体组织和卵巢滤泡细胞中表达。通过体外转录分析,我们鉴定出一个单一的12碱基对DNA元件,它能激活两个Yp基因启动子的转录。在体内,这个调控元件具有组织特异性:它能激活滤泡细胞中Yp1和Yp2报告基因的转录,但在脂肪体或其他组织中没有可检测到的作用。该元件的序列由转录因子GATA家族的两个识别位点组成。我们发现,在已知编码GATA因子的果蝇基因中,只有dGATAb在卵巢中表达。我们在卵巢中检测到的单一转录本经过可变剪接或起始,产生一种卵巢特异性的蛋白质异构体。细菌表达的dGATAb能与12碱基对元件结合;用于体外转录分析的Kc0核提取物中也存在类似的结合活性。这些体外和体内实验结果使我们提出,dGATAb产生几种受发育调控的产物,其中一种是滤泡细胞特异性蛋白,它能从一个已知的调控元件激活Yp1和Yp2的转录。