Hui C C, Suzuki Y
Department of Developmental Biology, National Institute for Basic Biology, Okazaki, Japan.
Gene. 1989 Dec 28;85(2):403-11. doi: 10.1016/0378-1119(89)90433-2.
The activity of the upstream promoter element (UPE) of the Bombyx mori fibroin-encoding gene was analyzed by using the major late promoter (MLP) of adenovirus 2 (Ad2) as the basal promoter. The UPE reveals remarkable stimulatory activity on the transcription of Ad2 MLP in tissue extracts prepared from the posterior silk gland where the fibroin-encoding gene is specifically expressed. This stimulation of transcription is due to an increase of initiation complex formation as shown by a one-round transcription assay. The UPE of the sericin-1-encoding gene, which is specifically expressed in the middle silk gland, also stimulates Ad2 MLP transcription in a similar manner. The results of the competition experiments indicate that the stimulatory effect is mediated by trans-acting factors which bind to these UPE. These two UPE are also active in tissue extracts of the middle silk gland, but are nearly inactive in extracts from the ovarian tissue and an embryonic cell line, Bm-e21. These observations further support the hypothesis that a silk-gland-specific factor(s) is responsible for this tissue-specific transcription in vitro. The possible molecular basis of this silk-gland specificity is discussed in light of the various trans-acting factors known to bind these UPE.
以腺病毒2(Ad2)的主要晚期启动子(MLP)作为基础启动子,对家蚕丝素编码基因的上游启动子元件(UPE)的活性进行了分析。在丝素编码基因特异性表达的后部丝腺制备的组织提取物中,UPE对Ad2 MLP的转录显示出显著的刺激活性。一轮转录分析表明,这种转录刺激是由于起始复合物形成的增加。在中部丝腺中特异性表达的丝胶蛋白1编码基因的UPE,也以类似的方式刺激Ad2 MLP转录。竞争实验结果表明,这种刺激作用是由与这些UPE结合的反式作用因子介导。这两个UPE在中部丝腺的组织提取物中也具有活性,但在卵巢组织和胚胎细胞系Bm-e21的提取物中几乎没有活性。这些观察结果进一步支持了这样的假设,即丝腺特异性因子负责体外这种组织特异性转录。根据已知与这些UPE结合的各种反式作用因子,讨论了这种丝腺特异性的可能分子基础。