Huber Wade E, Price E Roydon, Widlund Hans R, Du Jinyan, Davis Ian J, Wegner Michael, Fisher David E
Division of Pediatric Hematology/Oncology, Dana-Farber Cancer Institute and Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
J Biol Chem. 2003 Nov 14;278(46):45224-30. doi: 10.1074/jbc.M309036200. Epub 2003 Aug 27.
alpha-Melanocyte-stimulating hormone (MSH) utilizes cAMP to trigger pigmentation of melanocytes via activation of melanocyte-restricted microphthalmia-associated transcription factor (M-MITF) expression. M-MITF is a melanocyte-restricted helix-loop-helix transcription factor capable of transactivating promoters for multiple genes whose products modulate pigmentation. Although M-MITF promoter activation by MSH is known to occur through a conserved cAMP-response element (CRE), it remains unclear how this CRE exhibits such exquisitely tissue-restricted responsiveness. Here we show that cAMP-mediated CRE-binding protein activation of the M-MITF promoter requires a second DNA element located approximately 100 bp upstream, a site that is bound and activated by SOX10. Mutations in the SOX10 transcription factor, like MITF, results in a disorder known as Waardenburg Syndrome. The cAMP response of the M-MITF promoter was analyzed in melanoma and neuroblastoma cells (which are neural crest-derived but lack both M-MITF and SOX10 expression). M-MITF promoter responsiveness to cAMP was found to depend upon SOX10, and reciprocally, SOX10 transactivation was dependent upon the CRE. Ectopic SOX10 expression, in cooperation with cAMP signaling, activated the M-MITF promoter function and the expression of measurable endogenous M-MITF transcripts in neuroblastoma cells. SOX10dom, a mutant allele, failed to cooperate with cAMP in neuroblastoma cells and attenuated the cAMP responsiveness of the M-MITF promoter in melanoma cells. These observations demonstrate a means whereby the ubiquitous cAMP signaling machinery is harnessed to produce a highly tissue-restricted transcriptional response by cooperating with architectural factors, in this case SOX10.
α-黑素细胞刺激素(MSH)通过激活黑素细胞特异性小眼相关转录因子(M-MITF)的表达,利用环磷酸腺苷(cAMP)触发黑素细胞的色素沉着。M-MITF是一种黑素细胞特异性的螺旋-环-螺旋转录因子,能够反式激活多个基因的启动子,这些基因的产物可调节色素沉着。虽然已知MSH通过保守的环磷酸腺苷反应元件(CRE)激活M-MITF启动子,但目前尚不清楚该CRE如何表现出如此精确的组织限制性反应。在这里,我们表明,cAMP介导的CRE结合蛋白对M-MITF启动子的激活需要位于上游约100 bp处的第二个DNA元件,该位点由SOX10结合并激活。与MITF一样,SOX10转录因子的突变会导致一种名为Waardenburg综合征的疾病。在黑素瘤和神经母细胞瘤细胞(它们源自神经嵴,但缺乏M-MITF和SOX10表达)中分析了M-MITF启动子对cAMP的反应。发现M-MITF启动子对cAMP的反应性取决于SOX10,反之亦然,SOX10的反式激活取决于CRE。异位表达SOX10并与cAMP信号传导协同作用,可激活神经母细胞瘤细胞中的M-MITF启动子功能以及可测量的内源性M-MITF转录本的表达。突变等位基因SOX10dom在神经母细胞瘤细胞中无法与cAMP协同作用,并减弱了黑素瘤细胞中M-MITF启动子对cAMP的反应性。这些观察结果证明了一种机制,即通过与结构因子(在这种情况下为SOX10)合作,利用普遍存在的cAMP信号传导机制产生高度组织限制性的转录反应。