TRANSCRIPTIONAL REGULATION OF EPIDERMAL GENES

我的主要目标是阐明控制细胞的机制 角质形成细胞的命运和分化。 编码角蛋白的基因 K5和K14在细胞分裂过程中大量且独特地转录， 所有复层组织的角质形成细胞。因此，这些基因是理想的 探索顺式元件和转录因子的作用， 上皮特异性一些最小顺式作用元件 这些基因的启动子已经被鉴定，但是足够的区域 对于组织特异性和分化特异性基因表达， 尚未查明的上游分子。 使用DNA酶I超敏反应 通过分析，我现在已经发现了新的上游DNA序列， 染色质的细胞类型特异性变化。 为了测试这些区域 在角质细胞特异性基因表达中确实很重要，我将 a）确定角质形成细胞特异性蛋白质是否导致细胞凋亡， I观察到类型特异性DNA酶超敏反应，B）使用诱变， 角质形成细胞转染和转基因小鼠，以评估是否 区域在功能上是重要的，c）克隆和表征 控制K5和K14基因表达的转录因子，和d） 评估这些因素与角质形成细胞测定的相关性 细胞命运 这些知识也将是非常有价值的优化 角质形成细胞作为基因治疗和药物递送的载体。

My primary objective is to elucidate the mechanisms that control cell fate and differentiation in keratinocytes. The genes encoding keratins, K5 and K14 are abundantly and uniquely transcribed in dividing keratinocytes of all stratified tissues. Hence these genes are ideal to explore the role of cis-elements and transcription factors that confer epithelial-specificity. Some cis-acting elements of the minimal promoters of these genes have been identified, but regions sufficient for tissue specific and differentiation-specific gene expression include as yet unidentified upstream elements. Using DNAse I hypersensitivity assays, I have now uncovered novel upstream DNA sequences that display cell-type specific changes in chromatin. To test whether these regions are indeed important in keratinocyte-specific gene expression, I will a) determine whether keratinocyte-specific proteins account for cell type-specific DNAse hypersensitivity I observed, b) use mutagenesis, keratinocyte transfection and transgenic mice to assess whether the regions are functionally important, c) clone and characterize the transcription factors that govern K5 and K14 gene expression, and d) assess the relevance of these factors to determination of keratinocyte cell fate. This knowledge will also be extremely valuable in optimizing the keratinocyte as a vehicle for gene-therapy and drug delivery.