Zhukovskaya N, Early A, Kawata T, Abe T, Williams J
MRC Laboratory of Molecular Cell Biology, University College London, United Kingdom.
Dev Biol. 1996 Oct 10;179(1):27-40. doi: 10.1006/dbio.1996.0239.
In the Dictyostelium slug there are two types of prestalk cells, pstA cells and pstO cells, that differ in their ability to utilize the distal and proximal parts of the promoter of ecmA, a gene that is specifically expressed in prestalk cells. When Rm, a dominant inhibitory form of the regulatory subunit of cAMP-dependent protein kinase (PKA), is expressed under the control of the complete promoter of the ecmA gene (in a construct termed ecmAO:Rm) development proceeds to the slug stage. Although able to form small but outwardly normal slugs, ecmAO:Rm cells are defective in prestalk cell differentiation. In ecmAO:Rm cells, the induction of pstA- and pstO-specific gene expression by the stalk cell inducer DIF is greatly inhibited. Paradoxically, a very large fraction of the cells in an ecmAO:Rm slug show evidence of once having expressed the ecmA and ecmO prestalk markers. However, we present evidence that this is due to abortive prestalk cell differentiation that terminates when sufficient Rm protein has accumulated to block PKA activity. This results in regulative transdifferentiation of prespore cells to form prestalk cells. During their transitory period as prestalk cells the ecmAO:Rm cells coexpress both the ecmA and ecmO markers, indicating a possible link between PKA activity and divergence of the two prestalk cell subtypes. Finally, we show that the level of the DNA binding activity believed to lie at the end of the DIF signal transduction pathway is reduced in ecmAO:Rm slugs.
在盘基网柄菌蛞蝓体中有两种前柄细胞,即pstA细胞和pstO细胞,它们在利用ecmA基因启动子远端和近端区域的能力上存在差异,ecmA基因是一种在前柄细胞中特异性表达的基因。当cAMP依赖性蛋白激酶(PKA)调节亚基的显性抑制形式Rm在ecmA基因的完整启动子控制下表达时(在一个称为ecmA O:Rm的构建体中),发育会进行到蛞蝓体阶段。尽管ecmA O:Rm细胞能够形成小的但外观正常的蛞蝓体,但其在前柄细胞分化方面存在缺陷。在ecmA O:Rm细胞中,柄细胞诱导剂DIF对pstA和pstO特异性基因表达的诱导受到极大抑制。矛盾的是,ecmA O:Rm蛞蝓体中很大一部分细胞显示出曾经表达过ecmA和ecmO前柄标记物的证据。然而,我们提供的证据表明,这是由于前柄细胞分化失败所致,当积累了足够的Rm蛋白以阻断PKA活性时,这种分化就会终止。这导致前孢子细胞进行调节性转分化以形成前柄细胞。在作为前柄细胞的过渡阶段,ecmA O:Rm细胞同时共表达ecmA和ecmO标记物,这表明PKA活性与两种前柄细胞亚型的分化之间可能存在联系。最后,我们表明,在ecmA O:Rm蛞蝓体中,被认为位于DIF信号转导途径末端的DNA结合活性水平降低。