MOLECULAR BASIS FOR GATA-5 GENE EXPRESSION

GATA-5 基因表达的分子基础

• 批准号: 2445031
• 负责人: COLIN MACNEILL
• 金额: $ 8.53万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-05 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2445031
• 关键词: developmental genetics; enzyme induction /repression; gene expression; genetic promoter element; genetic regulatory element; genetically modified animals; heart; heart cell; histogenesis; laboratory mouse; mammalian embryology; myocardium; nucleic acid sequence; reporter genes; tissue /cell culture; transcription factor; transfection

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.

我完成了妇产科的临床培训， 获得为期三年的医生科学家奖，以追求熟练 在基础科学，与研究的最终职业目标，教学和 妇产科临床实践 这个奖项快结束了，我寻求两个 多年的MCSDA支持，以扩大我在分子生物学的熟练程度， 一个独立调查员。 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 一个新的三个加塔转录因子基因（加塔-4，加塔-4）的亚家族， 5，加塔-6），其在发育中的心脏中差异表达。 的 该基因的表达代表了最早的已知标记之一， 心前中胚层。 此外，加塔-5基因在大肠杆菌中得到了稳健表达 而加塔-4和加塔-6基因 在后半期的心脏中显著下调， 胚胎发育 我对启动子进行了测序， 基因内和基因侧翼的DNA酶I超敏感位点的数量。 我 建议对顺式作用元件和反式作用因子进行表征 调节加塔-5基因表达的基因。 我将确定控制区域 足以指导报告基因在心脏中特异性表达， 瞬时转基因小鼠。 然后，我将确定必要的方面， 这些区域在原发性心脏 肌细胞 我已经确定加塔-5基因在原发性 心肌细胞培养物，这些培养物适用于 瞬时转染测定。 确定的最低限度控制要素 将在转基因小鼠中进一步测试转染测定。 以来 加塔-5基因在内皮细胞和心肌细胞中表达， 细胞，这些研究还将揭示是否不同或共同控制 在这两个不同的层中， 的心脏。 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 赞助商约翰B博士。E.伯奇，是一个有成就的科学家， 转录调控领域。 他是福克斯的终身会员 大通癌症中心，这是一个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 科. 福克斯蔡斯癌症中心提供了一个杰出的学术 在基础和临床科学的培训环境。