PHOSPHORYLATION AND THE REGULATION OF RNA SPLICING

磷酸化和 RNA 剪接的调控

• 批准号: 6343045
• 负责人: WILLIAM W MATTOX
• 金额: $ 20.94万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6343045
• 关键词: Drosophilidae; RNA binding protein; RNA splicing; chemical binding; developmental genetics; gene expression; genetic mapping; genetic regulation; genetic strain; molecular assembly /self assembly; phenotype; phosphorylation; precursor mRNA; protein kinase; protein protein interaction; protein structure function; sex determination; sex differentiation; site directed mutagenesis; western blottings

Alternative pre-mRNA splicing affects the expression of gene products involved in many aspects of cell growth and differentiation, yet the mechanisms by which it is regulated are only beginning to be elucidated. SR proteins and related factors are thought to play important roles in the regulation of splicing. These factors form regulatory complexes at sites internal to the exons of alternatively spliced pre-mRNAs and function to recruit factors from the general splicing machinery to nearby splice sites that otherwise would not be recognized. Although SR proteins are highly phosphorylated, the role of phosphorylation in the regulation of splicing is largely unknown. We propose to investigate how SR protein phosphorylation affects the assembly and function of sex-specific regulatory complexes formed on a splicing enhancer in the doublesex pre-mRNA. Preliminary studies indicate that the LAMMER kinase Darkener-of- apricot phosphorylates SR proteins and is required for normal regulation of doublesex RNA splicing. We will test whether the phosphorylation of proteins from the doublesex splicing enhancer complex affects the ability of SR proteins, and related regulatory factors such as TRA and TRA2, to interact with each other and with the pre-mRNA. Other experiments will compare the effects of different classes of kinases on SR protein function. Finally, we will produce strains of Drosophila with mutations in SR protein kinases to examine their roles in development and on the phosphorylation of specific SR proteins in vivo. We anticipate that these studies will provide significant new insights into the mechanisms by which splicing is regulated and will open up new avenues for future studies on phosphorylation of SR proteins.

前体mRNA选择性剪接影响基因表达 产品涉及细胞生长的许多方面， 分化，但它的调节机制是 才刚刚开始被阐明。 SR蛋白及其相关因子 被认为在调节 拼接 这些因子在位点形成调节复合物 在选择性剪接的前mRNA的外显子内部， 从普通拼接机械中招募因子的功能 到附近的剪接位点，否则将无法识别。 尽管SR蛋白是高度磷酸化的，但 在剪接调节中的磷酸化在很大程度上是未知的。 我们建议研究SR蛋白磷酸化如何影响 性别特异性调节复合物的组装和功能 在doublemex前mRNA的剪接增强子上形成。 初步研究表明，LAMMER激酶 杏磷酸化SR蛋白质，是正常 doublemex RNA剪接的调节。 我们将测试 来自双链体前剪接增强子的蛋白质的磷酸化 复合物影响SR蛋白的能力， 调节因子，如TRA和TRA2，相互作用， 另一个和前mRNA。 其他实验将比较 不同种类的激酶对SR蛋白功能的影响。 最后，我们将生产出果蝇品系， SR蛋白激酶，以检查其在发育和 体内特异性SR蛋白的磷酸化。 我们 预计这些研究将提供重要的新的 深入了解剪接的调节机制， 将为今后的磷酸化研究开辟新的途径， SR蛋白。