MOLECULAR BASIS FOR GATA-5 GENE EXPRESSION

在完成产科/妇科临床培训后，我是 获得为期三年的内科科学家奖以追求熟练程度 在基础科学方面，最终的职业目标是研究、教学和 妇产科的临床实践。这个奖项快结束了，我想要两个 多年来对MCSDA的支持将我在分子生物学方面的熟练程度扩展到 一名独立调查员的身份。 该研究计划旨在提供有关 决定心脏发育的转录事件。我们已经描述了 一个新的三个GATA转录因子基因亚家族(GATA-4、GATA-4、GATA-2)。 5、GATA-6)，在发育心脏中差异表达。这个 该基因的表达代表了已知的最早的标记之一 心前中胚层。此外，GATA-5基因也得到了有力的表达。 而GATA-4和GATA-6基因 在下半个月心脏中的表达显著下调 胚胎发育。我已经对启动子进行了测序并绘制了一个 基因内部和侧翼的DNase I超敏感部位的数量。我 建议对顺式作用元件和反式作用因素进行表征 调控GATA-5基因表达的基因。我会确定控制区 足以指导心脏特异表达的报告基因在 瞬时转基因小鼠。然后，我将确定必要的方面 在原代心脏瞬时转染法中的这些区域 肌细胞。我已经确定gata-5基因在原代细胞中表达。 心肌细胞培养，这些培养物适合于 瞬时转染法。中确定的最小控制元素 将在转基因小鼠身上进行进一步的转基因测试。自.以来 GATA-5基因在心内膜细胞和心肌细胞中的表达 细胞，这些研究也将揭示不同的或共同的控制 该基因在这两个不同层面上的表达受多种因素控制。 心之所向。我相信这些实验将有助于 对我们理解心脏发育有重要意义。 发起人约翰·B·E·伯奇博士是世界上最有成就的科学家 转录调控域。他是福克斯的终身会员 大通癌症中心，NCI指定的综合癌症中心 ，宾夕法尼亚大学兼职副教授 医学院。他也是《分子》杂志的编辑委员会成员。 细胞生物学，是NIH生物科学2研究的成员 一节。福克斯·蔡斯癌症中心为一位杰出的学者提供 基础和临床科学训练的环境。

Having completed clinical training in obstetrics/gynecology, I am the recipient of a three year Physician Scientist Award to pursue proficiency in basic science, with the ultimate career goal of research, teaching and clinical practice of ob/gyn. This award is almost over, and I seek two years of MCSDA support to extend my proficiency in molecular biology to that of an independent investigator. The research plan is designed to provide information about the transcriptional events that dictate heart development. We have described a novel subfamily of three GATA transcription factor genes (gata-4,gata- 5,gata-6) that are differentially expressed in the developing heart. The expression of this gene represents one of the earliest known markers of pre-cardiac mesoderm. In addition, the gata-5 gene is robustly expressed in the heart throughout development whereas the gata-4 and gata-6 genes are dramatically downregulated in the heart during the latter half of embryonic development. I have sequenced the promoter and have mapped a number of DNase I hypersensitive sites within and flanking the gene. I propose to characterize the cis-acting elements and trans-acting factors that regulate gata-5 gene expression. I will identify control regions sufficient to direct heart-specific expression of a reporter gene in transient transgenic mice. I will then identify the necessary aspects of such regions in transient transfection assays in primary cardiac myocytes. I have determined that the gata-5 gene is expressed in primary cardiac myocyte cultures and that these cultures are suitable for transient transfection assays. Minimal control elements identified in transfection assays will be further tested in transgenic mice. Since the gata-5 gene is expressed in endocardial cells as well as myocardial cells, these studies will also reveal whether distinct or common control elements govern the expression of this gene in these two different layers of the heart. I believe that these experiments will contribute significantly to our understanding of cardiac development. The sponsor, Dr. John B. E. Burch, is an accomplished scientist in the field of transcription regulation. He is a tenured Member of the Fox Chase Cancer Center, which is an NCI designated Comprehensive Cancer Center, and an Adjunct Associate Professor at University of Pennsylvania Medical School. He is also on the editorial board of Molecular and Cellular Biology and is a member of the NIH Biological Sciences 2 Study Section. The Fox Chase Cancer Center affords an outstanding academic environment for training in basic and clinical science.