Biochemistry of lysine crotonylation pathway

赖氨酸巴豆酰化途径的生物化学

基本信息

批准号：
8654351
负责人：
YINGMING ZHAO
金额：
$ 42.37万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8654351
关键词：
Acetylation Acetyltransferase Acyltransferase Affinity Area Binding Proteins Biochemical Biochemistry Biological Biology Cell physiology Cells Chemicals Chromatin DNA Data Set Disease Drug Targeting Enhancers Enzymatic Biochemistry Enzymes Epigenetic Process Genes Genome Genomics Germ Cells Goals Haploid Cells Histone Acetylation Histones Human In Vitro Label Lead Libraries Link Lysine Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Meiosis Metals Methods Methylation Modification Mus Neurodegenerative Disorders Nuclear Extract Output Pathway interactions Peptide Library Peptides Phosphorylation Play Positioning Attribute Post-Translational Protein Processing Proteins Proteomics Recombinants Reporting Research Research Infrastructure Role Sex Chromosomes Sirtuins Site Spermatogenesis Structure Testing Testis Transcriptional Regulation Ubiquitination Western Blotting Work base diabetic enzyme substrate human disease in vivo insight male novel programs promoter protein complex protein structure function public health relevance response success

项目摘要

DESCRIPTION (provided by applicant): Post-translational modifications (PTMs) can significantly modulate the structure and function of proteins and are known to regulate a wide variety of cellular pathways. Dysregulation of PTM pathways have been shown to lead to diverse human diseases, such as cancer, diabetics, and neurodegenerative diseases. PTMs in histones, such as lysine acetylation (Kac) and lysine methylation (Kme), are major epigenetic marks, contributing to the epigenetic program that dictates diverse DNA-templated biological outputs and diseases. Enzymes that regulate histone PTMs other than phosphorylation (such as Kac, Kme, and lysine ubiquitination) have emerged as important drug targets for diseases. We recently reported the identification of a new, evolutionarily-conserved lysine modification, lysine crotonylation (Kcr). We showed that Kcr is abundant in core histones and identified 28 histone Kcr sites in mammalian cells. In male germinal cells immediately following meiosis, histone Kcr, but not histone Kac, is enriched on sex chromosomes and specifically marks testis-specific genes. The unique structure and genomic localization of histone Kcr suggest that it is dynamic and functionally different from histone Kac, a previously-described PTM with diverse functions. Despite of these preliminary studies, major players in the Kcr pathway remain unknown. We hypothesize that Kcr pathway has a unique set of regulatory enzymes, substrates and direct binders, which determines its difference from Kac. We therefore propose to use enzymology, chemical biology, and proteomics approaches to characterize Kcr pathway. Our team is well positioned to carry out this project because of our expertise and the tremendous relevant studies we have carried out in the past few years. In this project, we will first identify and characterize Kcr-regulatory enzymes, crotonyltransferase and decrotonylases, using a variety of chemical biology and enzymology approaches. We will then identify substrates of Kcr and study their dynamics in response to Kcr- regulatory enzymes and during spermatogenesis, by a quantitative proteomics approach. Finally, we will identify and confirm the direct binders for histone Kcr peptides. The new enzymes and binders for histone Kcr are likely to open entirely unforeseen fruitful avenues for histone biology and their roles in disease. Thus, the proposed study should accelerate current research in chromatin biology by revealing novel epigenetic mechanisms, perhaps analogous to the discovery of Kac- and Kme-regulatory enzymes for histone lysine acetylation and histone lysine methylation.

描述（由申请人提供）：翻译后修饰（PTM）可以显着调节蛋白质的结构和功能，并已知可以调节多种细胞途径。 PTM途径的失调已被证明会导致各种人类疾病，例如癌症，糖尿病患者和神经退行性疾病。组蛋白中的PTM，例如赖氨酸乙酰化（KAC）和赖氨酸甲基化（KME）是主要的表观遗传标记，这有助于表观遗传学程序，该计划决定了不同的DNA模拟生物产量和疾病。调节组蛋白PTM以外的其他磷酸化（例如KAC，KME和赖氨酸泛素化）的酶已成为疾病的重要药物靶标。我们最近报告了一种新的，进化保存的赖氨酸，赖氨酸的鉴定杂种（KCR）。我们表明，KCR在核心组蛋白中很丰富，并鉴定出哺乳动物细胞中的28个组蛋白KCR位点。在减数分裂后紧接的雄性生发细胞中，组蛋白KCR而不是组蛋白KAC富含性染色体，并特别标记睾丸特异性基因。组蛋白KCR的独特结构和基因组定位表明，它具有动态性，并且在功能上与组蛋白KAC（一种具有不同功能的PTM）的功能不同。尽管有这些初步研究，但KCR途径中的主要参与者仍然未知。我们假设KCR途径具有独特的调节酶，底物和直接粘合剂，这些酶决定了其与KAC的差异。因此，我们建议使用酶学，化学生物学和蛋白质组学方法来表征KCR途径。由于我们的专业知识以及过去几年中我们进行的巨大相关研究，我们的团队有能力执行该项目。在这个项目中，我们将使用各种化学生物学和酶学方法首先识别和表征KCR调节酶，Crotonyltransferase and Crotonyltransferase and Crotonylonylases。然后，我们将通过定量蛋白质组学方法来鉴定KCR的底物，并研究其对KCR调节酶以及精子发生过程中的动力学。最后，我们将识别并确认组蛋白KCR肽的直接粘合剂。组蛋白KCR的新酶和粘合剂可能会完全无法预料的组蛋白生物学及其在疾病中的作用。因此，拟议的研究应通过揭示新型的表观遗传机制来加速当前的染色质生物学研究，这可能类似于发现组蛋白碱性乙酰化和组蛋白赖氨酸赖氨酸甲基化的KAC-和KME调控酶。