Faure M, Franke J, Hall A L, Podgorski G J, Kessin R H
Department of Anatomy and Cell Biology, College of Physicians and Surgeons of Columbia University, New York, New York 10032.
Mol Cell Biol. 1990 May;10(5):1921-30. doi: 10.1128/mcb.10.5.1921-1930.1990.
The cyclic nucleotide phosphodiesterase (phosphodiesterase) plays essential roles throughout the development of Dictyostelium discoideum. It is crucial to cellular aggregation and to postaggregation morphogenesis. The phosphodiesterase gene is transcribed into three mRNAs, containing the same coding sequence connected to different 5' untranslated sequences, that accumulate at different times during the life cycle. A 1.9-kilobase (kb) mRNA is specific for growth, a 2.4-kb mRNA is specific for aggregation, and a 2.2-kb mRNA is specific for late development and is only expressed in prestalk cells. Hybridization of RNA isolated from cells at various stages of development with different upstream regions of the gene indicated separate promoters for each of the three mRNAs. The existence of specific promoters was confirmed by fusing the three putative promoter regions to the chloramphenicol acetyltransferase reporter gene, and the analysis of transformants containing these constructs. The three promoters are scattered within a 4.1-kilobase pair (kbp) region upstream of the initiation codon. The late promoter is proximal to the coding sequence, the growth-specific promoter has an initiation site that is 1.9 kbp upstream of the ATG codon, and the aggregation-specific promoter has an initiation site 3 kbp upstream.
环核苷酸磷酸二酯酶在盘基网柄菌的整个发育过程中发挥着重要作用。它对细胞聚集和聚集后形态发生至关重要。磷酸二酯酶基因转录为三种mRNA,它们含有连接到不同5'非翻译序列的相同编码序列,在生命周期的不同时间积累。一种1.9千碱基(kb)的mRNA在生长阶段特异表达,一种2.4 kb的mRNA在聚集阶段特异表达,一种2.2 kb的mRNA在发育后期特异表达且仅在前柄细胞中表达。用该基因不同上游区域与从不同发育阶段细胞中分离的RNA进行杂交,表明这三种mRNA各有独立的启动子。通过将三个推定的启动子区域与氯霉素乙酰转移酶报告基因融合,并对含有这些构建体的转化体进行分析,证实了特定启动子的存在。这三个启动子分散在起始密码子上游4.1千碱基对(kbp)的区域内。后期启动子靠近编码序列,生长特异性启动子的起始位点在ATG密码子上游1.9 kbp处,聚集特异性启动子的起始位点在ATG密码子上游3 kbp处。