{"title":"在10T1/2成纤维细胞中,MyoD对心肌MLC-2启动子的转激活与E-box的要求无关,但取决于识别MEF-2位点的新蛋白。","authors":"S K Goswami, M A Siddiqui","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>MyoD-mediated activation of skeletal muscle genes, which is dependent upon the consensus E-box sequence, involves, at least in one group of muscle genes, another transcription factor, the myocyte enhancer factor-2 (MEF-2). Since the cardiac myosin light chain-2 (MLC-2) gene promoter lacks the functional E-box but contains the activator MEF-2 site, we tested the effect of ectopic expression of MyoD on cardiac MLC-2 promoter function. Here, we demonstrate that either transient or stable expression of MyoD in otherwise nonpermissive C3H10T1/2 fibroblast cells can promote the expression of MLC-2/CAT. Deletion and site-specific mutation analysis demonstrate that the MEF-2 site (Element B) in the MLC-2 promoter is the target of activation by MyoD. Gel mobility shift assay using nuclear extracts from the normal and MyoD-transfected fibroblast cells did not show a difference in the major MEF-2 binding complexes, except for one complex of fast-moving mobility, which suggested that new MEF-2-like regulatory proteins are induced by MyoD.</p>","PeriodicalId":72545,"journal":{"name":"Cellular & molecular biology research","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Transactivation of cardiac MLC-2 promoter by MyoD in 10T1/2 fibroblast cells is independent of E-box requirement but depends upon new proteins that recognize MEF-2 site.\",\"authors\":\"S K Goswami, M A Siddiqui\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>MyoD-mediated activation of skeletal muscle genes, which is dependent upon the consensus E-box sequence, involves, at least in one group of muscle genes, another transcription factor, the myocyte enhancer factor-2 (MEF-2). Since the cardiac myosin light chain-2 (MLC-2) gene promoter lacks the functional E-box but contains the activator MEF-2 site, we tested the effect of ectopic expression of MyoD on cardiac MLC-2 promoter function. Here, we demonstrate that either transient or stable expression of MyoD in otherwise nonpermissive C3H10T1/2 fibroblast cells can promote the expression of MLC-2/CAT. Deletion and site-specific mutation analysis demonstrate that the MEF-2 site (Element B) in the MLC-2 promoter is the target of activation by MyoD. Gel mobility shift assay using nuclear extracts from the normal and MyoD-transfected fibroblast cells did not show a difference in the major MEF-2 binding complexes, except for one complex of fast-moving mobility, which suggested that new MEF-2-like regulatory proteins are induced by MyoD.</p>\",\"PeriodicalId\":72545,\"journal\":{\"name\":\"Cellular & molecular biology research\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1995-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cellular & molecular biology research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cellular & molecular biology research","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Transactivation of cardiac MLC-2 promoter by MyoD in 10T1/2 fibroblast cells is independent of E-box requirement but depends upon new proteins that recognize MEF-2 site.
MyoD-mediated activation of skeletal muscle genes, which is dependent upon the consensus E-box sequence, involves, at least in one group of muscle genes, another transcription factor, the myocyte enhancer factor-2 (MEF-2). Since the cardiac myosin light chain-2 (MLC-2) gene promoter lacks the functional E-box but contains the activator MEF-2 site, we tested the effect of ectopic expression of MyoD on cardiac MLC-2 promoter function. Here, we demonstrate that either transient or stable expression of MyoD in otherwise nonpermissive C3H10T1/2 fibroblast cells can promote the expression of MLC-2/CAT. Deletion and site-specific mutation analysis demonstrate that the MEF-2 site (Element B) in the MLC-2 promoter is the target of activation by MyoD. Gel mobility shift assay using nuclear extracts from the normal and MyoD-transfected fibroblast cells did not show a difference in the major MEF-2 binding complexes, except for one complex of fast-moving mobility, which suggested that new MEF-2-like regulatory proteins are induced by MyoD.