ALTERNATIVE SPLICING OF CONTRACTILE PROTEIN GENES

描述：（改编自研究者摘要）：总体目标 在本申请中提出的研究仍然是 顺式和反式作用因子鉴定和表征 参与了可变剪接的调节。 为此中央 研究人员将继续使用微型基因构建体， 来自收缩蛋白基因α-原肌球蛋白和 肌钙蛋白T作为模型系统，将使用体内和体内 体外无细胞剪接系统。 将追求三个主要具体目标： 1.-分析和表征的结构和作用机制， 剪接因子，聚嘧啶结合蛋白（PBP），他们有 最近被提纯并部分测序 这个因素起着重要的作用。 在决定3'剪接强度中的作用，并可能参与早期 剪接位点提交的步骤。 2.-为了进一步确定结合位点 平滑肌细胞特有的负剪接因子 为了隔离， 纯化，并描述该因子的作用机制， 通过阻止默认的拼接模式。 3.-使用高度选择性的 遗传学方法，鉴定和表征阳性剪接因子 特异于成熟平滑肌细胞，其决定调节的 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 拼接因子将对理解 剪接的组成性和选择性模型。

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.