The role of the cyclic nucleotide phosphodiesterase of Dictyostelium discoideum during growth, aggregation, and morphogenesis: overexpression and localization studies with the separate promoters of the pde.
The role of the cyclic nucleotide phosphodiesterase of Dictyostelium discoideum during growth, aggregation, and morphogenesis: overexpression and localization studies with the separate promoters of the pde.
复制标题
盘基网柄菌环核苷酸磷酸二酯酶在生长、聚集和形态发生过程中的作用：使用 pde 的单独启动子进行过表达和定位研究。
DOI：
10.1006/dbio.1993.1113
复制
发表时间：
1993
影响因子：
2.7
通讯作者：
Kessin,RH
中科院分区：
文献类型：
--
作者：
Hall,AL;Franke,J;Faure,M;Kessin,RH
在盘基网柄藻的整个发育过程中，cAMP作为一种初级信号，受一种分泌的环核苷酸磷酸二酯酶（PDE）的调节。PDE基因（pde）的表达由营养生长（prV）、聚集（prA）或晚期发育（prL）特异性启动子控制。将含有启动子的区域单独地融合到pdecoding序列。转化后，每个构建体的多个拷贝导致PDE mRNA和酶活性的过表达，其中每个启动子具有预期的时间分布。与对照转化体相比，来自PRV和PrA的PDE的过表达改变了聚集的时间，但最终形态是正常的。对照转化体与非转化细胞相比显示出延迟的聚集。从prL过表达PDE的细胞像对照转化体一样聚集，但没有形成子实体。单个启动子区与β-半乳糖苷酶基因（lacZ）融合。表达prA-lacZ的细胞分散在聚集区和土丘中。首先看到表达prL-lacZ的细胞均匀地分布在整个紧密和尖端土丘。在蛞蝓中，这些细胞大多位于前部区域。在顶点期间，表达prL-lacZ构建体的细胞被并入柄中，并且在孢子团周围的上部和下部杯中可见。
cAMP acts as a primary signal and is regulated by a secreted cyclic nucleotide phosphodiesterase (PDE) throughout development inDictyostelium discoideum. Expression of the PDE gene (pde) is controlled by promoters specific to vegetative growth (prV), aggregation (prA), or late development (prL). Promoter-containing regions were individually fused to thepdecoding sequence. After transformation multiple copies of each construct led to overexpression of PDE mRNA and enzyme activity with the temporal profile expected of each promoter. Overexpression of PDE from prV and prA altered the timing of aggregation compared to control transformants, but the final morphology was normal. Control transformants showed delayed aggregation compared to nontransformed cells. Cells that overexpressed PDE from prL aggregated like the control transformants, but no fruiting bodies were formed. Individual promoter regions were fused to the β-galactosidase gene (lacZ). Cells that expressed prA-lacZ were dispersed throughout aggregation fields and mounds. Cells that expressed prL-lacZ were first seen distributed homogeneously throughout tight and tipped mounds. In slugs most of these cells are localized in the anterior region. During culmination, cells that expressed the prL-lacZ construct became incorporated into the stalk and were seen in the upper and lower cups surrounding the spore mass.