The Functional Analysis of the Coactivator CARM1

描述(申请人提供)：精氨酸甲基化是一种广泛存在于核蛋白和细胞质蛋白上的翻译后修饰。精氨酸残基的甲基化是由蛋白质精氨酸N-甲基转移酶(PRMT)家族催化的，在哺乳动物中至少有9个成员。PRMT在进化上是保守的，在酵母和人类之间都有发现，但在细菌中没有。精氨酸甲基化的生物学后果在很大程度上是未知的，尽管随着特定底物的鉴定和最近小鼠模型的发展，功能正在显现。我们的目标是阐明共激活因子相关精氨酸甲基转移酶1(CARM1)的生物学作用，它是转录辅助激活因子。CARM1如何作为辅助激活因子发挥作用尚不清楚。它甲基化组蛋白H3、p300、PABP1和一组剪接因子。我们计划鉴定和鉴定被招募到CARM1甲基化基序的蛋白质，并评估CARM1在剪接中的作用。我们已经产生了CARM1基因敲除小鼠，来自突变胚胎的细胞将为体内分析假定的底物、甲基依赖结合蛋白和剪接研究提供一个基因控制的工具。Knockin CARM1小鼠也将被产生和分析，以评估这种分子的酶活性的重要性，而不是它的支架功能。最终，对CARM1功能的深入分析将导致对一类新的非调控酶的机制理解，这些酶在许多疾病状态中发挥作用，并成为药物开发的新靶点。

DESCRIPTION (provided by applicant): Arginine methylation is a widespread posttranslational modification found on both nuclear and cytoplasmic proteins. The methylation of arginine residues is catalyzed by the protein arginine N-methyltransferase (PRMT) family of enzymes, of which there are at least nine members in mammals. PRMTs are evolutionarily conserved and are found from yeast to man, but not in bacteria. The biological consequences of arginine methylation are largely unknown, although with the identification of specific substrates and the recent development of mouse models, functions are emerging. Our goal is to elucidate the biological role of coactivator-associated arginine methyltransferase1 (CARM1), which is transcriptional coactivator. How CARM1 functions as a coactivator is unclear. It methylates histone H3, p300, PABP1 and a subset of splicing factors. We plan to identify and characterize proteins that are recruited to the CARM1 methylated motifs and evaluate the role of CARM1 in splicing. We have generated CARM1 knockout mice, and cells derived from mutant embryos will provide a genetically controlled tool for the in vivo analysis of putative substrates, methyl-dependent binding proteins and splicing studies. Knockin CARM1 mice will also be generated and analyzed to evaluate the importance of this molecules enzymatic activity as opposed to its scaffolding functions. Ultimately, this in-depth analysis of CARM1 function will lead to the mechanistic understanding of a novel class of deregulated enzymes that play a role in a number of disease states and are new targets for drug development.