Escalante R, Sastre L
Instituto de Investigaciones Biomédicas del Consejo Superior de Investigaciones Científicas, C/Arturo Duperier, 4, 28029 Madrid, Spain.
Biochem Biophys Res Commun. 2001 Jul 20;285(3):820-4. doi: 10.1006/bbrc.2001.5235.
The MADS-box-containing gene srfA from Dictyostelium discoideum codes for a putative transcription factor that plays multiple roles in the development of this social amoeba. We have investigated the regulation of srfA gene expression after disaggregation of the cells from developing structures. The steady-state level of srfA mRNA was strongly and transiently induced shortly after disaggregation. srfA is maximally expressed 20 min after cell disaggregation and decreases thereafter. Induction was not dependent on protein synthesis, PKA, the kinase SplA and SrfA itself. This phenomena does not occur when cells are disaggregated in a small volume of buffer, suggesting the presence of extracellular molecules that repress srfA gene expression. To test this hypothesis, several well-known extracellular signaling molecules were studied. We found that srfA mRNA induction can be efficiently repressed by addition of exogenous cAMP and DIF-1 to the buffer in which the cells were disaggregated. Addition of other extracellular compounds such as ammonia, adenosine, SDF-1, and SDF-2 had no effect. srfA promoter P2, specifically induced during slug migration, was responsible for this regulation by extracellular compounds.
盘基网柄菌中含MADS盒的基因srfA编码一种假定的转录因子,该转录因子在这种社会性变形虫的发育中发挥多种作用。我们研究了从发育结构中解离细胞后srfA基因表达的调控情况。解离后不久,srfA mRNA的稳态水平被强烈且短暂地诱导。srfA在细胞解离后20分钟时表达量最高,此后下降。诱导不依赖于蛋白质合成、蛋白激酶A、激酶SplA和SrfA本身。当细胞在少量缓冲液中解离时,这种现象不会发生,这表明存在抑制srfA基因表达的细胞外分子。为了验证这一假设,我们研究了几种著名的细胞外信号分子。我们发现,通过向细胞解离所用的缓冲液中添加外源性环磷酸腺苷(cAMP)和分化诱导因子1(DIF-1),可以有效抑制srfA mRNA的诱导。添加其他细胞外化合物,如氨、腺苷、基质细胞衍生因子1(SDF-1)和基质细胞衍生因子2(SDF-2),则没有效果。在蛞蝓迁移过程中特异性诱导的srfA启动子P2负责这种由细胞外化合物介导的调控。