Spencer J A, Major M L, Misra R P
Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
Mol Cell Biol. 1999 Jun;19(6):3977-88. doi: 10.1128/MCB.19.6.3977.
Serum response factor (SRF) plays a central role in the transcriptional response of mammalian cells to a variety of extracellular signals. It is a key regulator of many cellular early response genes which are believed to be involved in cell growth and differentiation. The mechanism by which SRF activates transcription in response to mitogenic agents has been extensively studied; however, significantly less is known about regulation of the SRF gene itself. Previously, we identified distinct regulatory elements in the SRF promoter that play a role in activation, including a consensus ETS domain binding site, a consensus overlapping Sp/Egr-1 binding site, and two SRF binding sites. We further showed that serum induces SRF by a mechanism that requires an intact SRF binding site, also termed a CArG box. In the present study we demonstrate that in response to stimulation of cells by a purified growth factor, basic fibroblast growth factor (bFGF), the SRF promoter is upregulated by a complex pathway that involves at least two independent mechanisms: a CArG box-independent mechanism that is mediated by an ETS binding site, and a novel CArG box-dependent mechanism that requires both an Sp factor binding site and the CArG motifs for maximal stimulation. Our analysis indicates that the CArG/Sp element activation mechanism is mediated by distinct signaling pathways. The CArG box-dependent component is targeted by a Rho-mediated pathway, and the Sp binding site-dependent component is targeted by a Ras-mediated pathway. Both SRF and bFGF have been implicated in playing an important role in mediating cardiogenesis during development. The implications of our findings for SRF expression during development are discussed.
血清反应因子(SRF)在哺乳动物细胞对多种细胞外信号的转录反应中起核心作用。它是许多细胞早期反应基因的关键调节因子,这些基因被认为参与细胞生长和分化。SRF响应有丝分裂原激活转录的机制已得到广泛研究;然而,关于SRF基因本身的调控却知之甚少。此前,我们在SRF启动子中鉴定出了在激活过程中起作用的不同调控元件,包括一个共有ETS结构域结合位点、一个共有重叠Sp/Egr-1结合位点和两个SRF结合位点。我们进一步表明,血清通过一种需要完整SRF结合位点(也称为CArG盒)的机制诱导SRF。在本研究中,我们证明,响应纯化的生长因子碱性成纤维细胞生长因子(bFGF)对细胞的刺激,SRF启动子通过一条复杂的途径上调,该途径至少涉及两种独立机制:一种由ETS结合位点介导的不依赖CArG盒的机制,以及一种新的依赖CArG盒的机制,该机制需要一个Sp因子结合位点和CArG基序才能实现最大刺激。我们的分析表明,CArG/Sp元件激活机制由不同的信号通路介导。依赖CArG盒的成分由Rho介导的途径靶向,依赖Sp结合位点的成分由Ras介导的途径靶向。SRF和bFGF都被认为在发育过程中介导心脏发生中起重要作用。我们讨论了这些发现对发育过程中SRF表达的影响。