Ginsburg G, Kimmel A R
Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9332-6. doi: 10.1073/pnas.86.23.9332.
We have previously shown that several genes expressed during Dictyostelium development could be induced in shaking culture by exogenous cAMP, even though the accumulation of intracellular cAMP was inhibited. The use of selected cAMP analogs indicated that the exogenous cAMP functioned by activating the cell surface cAMP receptor and not by interacting with the regulatory subunit of the intracellular cAMP-dependent protein kinase. Although some genes in Dictyostelium appear to be regulated by intracellular cAMP, these data suggest that this is not the case for all genes regulated by cAMP. Intracellular second messengers other than cAMP may, therefore, promote the expression of these other genes. Here, we have examined inositol trisphosphate and diacylglycerol as candidates for such mediators of signal transduction. We have studied three genes that exhibit disparate modes of temporal and spatial expression during development of Dictyostelium. In shaking cultures, maximal levels of expression of each are dependent on the accumulation of or exposure to extracellular cAMP. We show that the addition of inositol trisphosphate and/or diacylglycerol to cells in shaking culture has distinct effects on the expression of each gene and, under specific conditions, can bypass the requirement for extracellular cAMP. These data suggest that extracellular cAMP interacting with its cell surface receptor may promote synthesis of inositol trisphosphate and diacylglycerol to regulate gene expression and aspects of differentiation in Dictyostelium.
我们之前已经表明,尽管细胞内cAMP的积累受到抑制,但在盘基网柄菌发育过程中表达的几个基因可以在外源cAMP的作用下在振荡培养中被诱导。对选定的cAMP类似物的使用表明,外源cAMP通过激活细胞表面的cAMP受体起作用,而不是通过与细胞内cAMP依赖性蛋白激酶的调节亚基相互作用。虽然盘基网柄菌中的一些基因似乎受细胞内cAMP的调控,但这些数据表明并非所有受cAMP调控的基因都是如此。因此,除cAMP之外的细胞内第二信使可能促进这些其他基因的表达。在这里,我们研究了肌醇三磷酸和二酰基甘油作为这种信号转导介质的候选物。我们研究了在盘基网柄菌发育过程中表现出不同时空表达模式的三个基因。在振荡培养中,每个基因的最大表达水平取决于细胞外cAMP的积累或暴露。我们表明,向振荡培养的细胞中添加肌醇三磷酸和/或二酰基甘油对每个基因的表达有不同的影响,并且在特定条件下,可以绕过对细胞外cAMP的需求。这些数据表明,与细胞表面受体相互作用的细胞外cAMP可能促进肌醇三磷酸和二酰基甘油的合成,以调节盘基网柄菌中的基因表达和分化过程。