Lindsley Andrew, Snider Paige, Zhou Hongming, Rogers Rhonda, Wang Jian, Olaopa Michael, Kruzynska-Frejtag Agnieszka, Koushik Shrinagesh V, Lilly Brenda, Burch John B E, Firulli Anthony B, Conway Simon J
Cardiovascular Development Group, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Dev Biol. 2007 Jul 15;307(2):340-55. doi: 10.1016/j.ydbio.2007.04.041. Epub 2007 May 3.
Periostin is a fasciclin-containing adhesive glycoprotein that facilitates the migration and differentiation of cells that have undergone epithelial-mesenchymal transformation during embryogenesis and in pathological conditions. Despite the importance of post-transformational differentiation as a general developmental mechanism, little is known how periostin's embryonic expression is regulated. To help resolve this deficiency, a 3.9-kb periostin proximal promoter was isolated and shown to drive tissue-specific expression in the neural crest-derived Schwann cell lineage and in a subpopulation of periostin-expressing cells in the cardiac outflow tract endocardial cushions. In order to identify the enhancer and associated DNA binding factor(s) responsible, in vitro promoter dissection was undertaken in a Schwannoma line. Ultimately a 304-bp(peri) enhancer was identified and shown to be capable of recapitulating 3.9 kb(peri-lacZ)in vivo spatiotemporal patterns. Further mutational and EMSA analysis helped identify a minimal 37-bp region that is bound by the YY1 transcription factor. The 37-bp enhancer was subsequently shown to be essential for in vivo 3.9 kb(peri-lacZ) promoter activity. Taken together, these studies identify an evolutionary-conserved YY1-binding 37-bp region within a 304-bp periostin core enhancer that is capable of regulating simultaneous novel tissue-specific periostin expression in the cardiac outflow-tract cushion mesenchyme and Schwann cell lineages.
骨膜蛋白是一种含成束蛋白的黏附糖蛋白,在胚胎发育和病理状态下,它能促进经历上皮-间充质转化的细胞迁移和分化。尽管转化后分化作为一种普遍的发育机制很重要,但关于骨膜蛋白胚胎期表达的调控却知之甚少。为了弥补这一不足,研究人员分离出一个3.9kb的骨膜蛋白近端启动子,并发现它能驱动神经嵴来源的施万细胞系以及心脏流出道心内膜垫中表达骨膜蛋白的细胞亚群中的组织特异性表达。为了确定负责的增强子和相关DNA结合因子,研究人员在一个神经鞘瘤细胞系中进行了体外启动子剖析。最终,一个304bp的(peri)增强子被鉴定出来,并显示其能够在体内重现3.9kb(peri-lacZ)的时空模式。进一步的突变和电泳迁移率变动分析有助于确定一个由YY1转录因子结合的最小37bp区域。随后发现,这个37bp的增强子对于体内3.9kb(peri-lacZ)启动子活性至关重要。综上所述,这些研究在一个304bp的骨膜蛋白核心增强子中鉴定出一个进化保守的、与YY1结合的37bp区域,该区域能够调控心脏流出道垫间充质和施万细胞系中同时出现的新型组织特异性骨膜蛋白表达。