Telokin gene regulation in smooth muscle

描述(由申请人提供)：我们的长期目标是利用 以端粒酶启动子为模型系统确定调控机制 平滑肌特异性基因表达。解开这些机制是至关重要的 对于我们对平滑肌发育和许多 不同的血管、肺、肠道和泌尿生殖系统疾病 与收缩蛋白表达改变和改变有关 伸缩性。实验被用来检验模数的假设 转录元件控制不同血管中基因的表达 纸巾。转基因小鼠将被用来鉴定最小的细胞特异性 Telokin推动者。体外鉴定的重要顺式作用调控元件， 将以突变分析为特征，端粒蛋白启动子- 转基因β-半乳糖苷酶。以确定监管要素是否 对于基础表达或平滑肌特异性，我们将采用 使用端粒酶/SM22a嵌合启动子的新方法。 调节端粒蛋白启动子活性的几种转录因子有 已查明，包括SRF。HFH-L、HFH-8、CDP、TEF和HMGI。一个主要目标 这项建议的目的是确定这些因素在监管中发挥的作用 端粒蛋白等平滑肌收缩蛋白在正常人群中的表达 以及受损的表型修饰的平滑肌细胞。具体来说，我们将 验证hFH-L和顺铂抑制和hFH-8和TEF刺激的假说 端粒酶基因的表达。建筑转录因子的作用 HMGI(Y)对端粒酶启动子活性和收缩蛋白的调节作用 表达式也将被求值。众所周知，SRF是所有 到目前为止，已经检测到了平滑肌基因，尽管是否如此尚不清楚 需要的基础表达的平滑肌基因或如果它参与了 调节它们的组织特异性表达。很可能这些组织 SRF的特殊功能源于其与其他细胞类型的相互作用 因此，提出了改良酵母-2杂交筛选的具体因素 确定新的平滑肌限制性结合伙伴。初步结果 已经产生了两种有趣的蛋白质，UBC9和Duplin，尽管广泛地 表示，可能通过相扑对SRF的活动产生根本性影响 分别与β-连环蛋白的修饰和相互作用。的功能 SRF与这些蛋白质的相互作用将被阐明。

DESCRIPTION (provided by applicant): Our long-term goal is to utilize the telokin promoter as a model system to determine the mechanisms regulating smooth muscle-specific gene expression. Unraveling these mechanisms is crucial to our understanding of smooth muscle development and of the pathology of many different vascular, pulmonary, intestinal and urogenital diseases that are associated with altered contractile protein expression and altered contractility. Experiments are proposed to test the hypothesis that modular transcription elements control gene expression in different smooth muscle tissues. Transgenic mice will be utilized to identify the minimal cell-specific telokin promoter. Important cis-acting regulatory elements identified in vitro, will be characterized by mutational analysis, of telokin promoter- beta-galactosidase transgenes. To determine if regulatory elements are important for basal expression or smooth muscle specificity we will employ a novel approach using chimeric telokin/SM22a promoters. Several transcription factors that regulate the telokin promoter activity have been identified, including SRF. HFH-l, HFH-8, CDP, TEF and HMGI. A major goal of this proposal is to determine the role played by these factors in regulating expression of telokin and other smooth muscle contractile proteins in normal and injured, phenotypically modified, smooth muscle cells. Specifically we will test the hypothesis that HFH-l and CDP repress and HFH-8 and TEF stimulate telokin gene expression. The role of architectural transcription factors HMGI(Y) in regulating telokin promoter activity and contractile protein expression will also be evaluated. SRF is known to be a key regulator of all smooth muscle genes thus far examined, although it is not known if it is required for basal expression of smooth muscle genes or if it is involved in mediating their tissue specific expression. It is likely that the tissue specific functions of SRF result from its interaction with other cell-type specific factors, hence, modified yeast-2 hybrid screens are proposed to identify novel smooth muscle restricted binding partners. Preliminary results have yielded two interesting proteins, UBC9 and Duplin, which although widely expressed, may have fundamental effects on SRF's activity through Sumo modification and interaction with beta--catenin, respectively. The function of SRFs' interaction with these proteins will be elucidated.