Driscoll D M, Williams J G
Imperial Cancer Research Fund, Clare Hall Laboratories, Hertfordshire, United Kingdom.
Mol Cell Biol. 1987 Dec;7(12):4482-9. doi: 10.1128/mcb.7.12.4482-4489.1987.
The cysteine proteinase 1 (CP1) gene of Dictyostelium discoideum encodes a developmentally regulated sulfhydryl proteinase. We characterized the DNA sequences upstream of the CP1 gene and found a second developmentally regulated gene, which we term DG17. The translational open reading frame of the DG17 gene encoded a 458-amino-acid cysteine- and lysine-rich protein of unknown function. In several regions, the cysteine and lysine residues were arranged in a manner characteristic of the zinc-binding domains found in proteins which interact with nucleic acids. During normal development, the DG17 and CP1 genes are coordinately activated late in aggregation. The addition of exogenous cyclic AMP (cAMP) induced the premature expression of both mRNAs. By measuring the rate of specific mRNA synthesis in isolated nuclei, we showed that cAMP acted at the transcriptional level to activate both genes. The two genes were separated by 910 nucleotides and were divergently transcribed. The intergenic region was predominantly composed of A + T residues except for four short G-rich regions. These sequences coincided with the positions of four nuclease-hypersensitive sites, which appear during aggregation when the DG17 and CP1 genes are transcribed (J. Pavlovic, E. Banz, and R. W. Parish, Nucleic Acids Res. 14:8703-8722, 1986). Two of the G-rich regions formed the core of two almost identical 80-nucleotide repeats located 220 and 320 nucleotides upstream of the CP1 gene. Using the Dictyostelium transformation system, we showed that a restriction fragment containing the intergenic region was capable of directing bidirectional transcription in a cAMP-dependent manner.
盘基网柄菌的半胱氨酸蛋白酶1(CP1)基因编码一种受发育调控的巯基蛋白酶。我们对CP1基因上游的DNA序列进行了特征分析,发现了另一个受发育调控的基因,我们将其命名为DG17。DG17基因的翻译开放阅读框编码一个458个氨基酸的富含半胱氨酸和赖氨酸的蛋白质,其功能未知。在几个区域,半胱氨酸和赖氨酸残基的排列方式具有与与核酸相互作用的蛋白质中发现的锌结合结构域的特征。在正常发育过程中,DG17和CP1基因在聚集后期被协同激活。添加外源环磷酸腺苷(cAMP)可诱导两种mRNA的过早表达。通过测量分离细胞核中特异性mRNA合成的速率,我们表明cAMP在转录水平上起作用以激活这两个基因。这两个基因相隔910个核苷酸,呈反向转录。基因间区域除了四个短的富含G的区域外,主要由A + T残基组成。这些序列与四个核酸酶超敏位点的位置一致,当DG17和CP1基因转录时,这些位点在聚集过程中出现(J. Pavlovic、E. Banz和R. W. Parish,《核酸研究》14:8703 - 8722,1986)。其中两个富含G的区域形成了位于CP1基因上游220和320个核苷酸处的两个几乎相同的80个核苷酸重复序列的核心。使用盘基网柄菌转化系统,我们表明包含基因间区域的一个限制片段能够以cAMP依赖的方式指导双向转录。