ALTERNATIVE SPLICING OF CONTRACTILE PROTEIN GENES

描述：(改编自《调查员摘要》)：总体目标 在本申请中提出的研究的部分仍然是 顺式和反式作用因子的鉴定与鉴定 参与了选择性剪接的调控。为此， 研究人员将继续使用微型基因结构和特定的 收缩蛋白基因α-原肌球蛋白和 肌钙蛋白T作为一种模型系统，将在体内和在 体外无细胞剪接系统。主要的具体目标有三个： 1.-分析和表征一种 剪接因子，多嘧啶结合蛋白(PBP)，他们有 最近进行了纯化和部分测序。这个因素起着重要的作用。 在决定3‘端剪接强度中的作用，并可能参与早期 拼接现场承诺的步骤。2.-进一步确定结合部位 一种特定于平滑肌细胞的负剪接因子。为了隔离， 提纯并表征起作用的这一因子的作用机制 通过阻止默认的拼接模式。3.-使用高度选择性的 用遗传方法鉴定和鉴定正剪接因子 对成熟的平滑肌细胞的特异性决定了其调控 肌钙蛋白T转录本的剪接模式。 预计这三个基因的克隆和鉴定 剪接因子将对理解 剪接的构造性和替代性模型。

DESCRIPTION: (Adapted from the Investigator's abstract): The overall goal of the research proposed in this application continues to be the identification and characterization of cis- and trans-acting factors involved in the regulation of alternative splicing. To this end, the investigators will continue to use mini-gene constructs and particular sequence elements from the contractile protein genes alpha-tropomyosin and troponin T as a model system that will be analyzed using in vivo and in vitro cell-free splicing system. Three main specific aims will be pursued: 1.-To analyze and characterize the structure and mechanism of action of a splicing factor, Polypyrimidine Binding Protein (PBP), which they have recently purified and partially sequenced. This factor plays an important role in determining 3' splice strength and might be involved in the early steps of splice site commitment. 2.-To further determine the binding site of a negative splicing factor specific to smooth muscle cells. To isolate, purify, and characterize the mechanism of action of this factor that works by blocking the default splicing pattern. 3.-Using a highly selective genetic approach, to identify and characterize a positive splicing factor specific to mature smooth muscle cells which determines the regulated splicing pattern of the troponin T transcript. It is expected that the cloning and characterization of these three splicing factors will make a significant contribution to the understanding of both constitutive and alternative models of splicing.