Biochemistry of lysine crotonylation pathway

赖氨酸巴豆酰化途径的生物化学

• 批准号: 8482624
• 负责人: YINGMING ZHAO
• 金额: $ 43.66万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 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/8482624
• 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

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）可显著调节蛋白质的结构和功能，已知可调节多种细胞途径。PTM通路的失调已显示导致多种人类疾病，如癌症、糖尿病和神经退行性疾病。组蛋白中的PTM，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，有助于决定不同DNA模板生物输出和疾病的表观遗传程序。调节组蛋白PTM而不是磷酸化的酶（如Kac、Kme和赖氨酸泛素化）已经成为疾病的重要药物靶标。我们最近报道了一种新的，进化上保守的赖氨酸修饰， 巴豆酰化（Kcr）。我们发现，Kcr是丰富的核心组蛋白，并确定了28组蛋白Kcr位点在哺乳动物细胞。在减数分裂后的雄性生殖细胞中，组蛋白Kcr（而不是组蛋白Kac）在性染色体上富集，并特异性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是动态的，并且在功能上不同于组蛋白Kac，组蛋白Kac是先前描述的具有不同功能的PTM。尽管有这些初步研究，Kcr途径的主要参与者仍然未知。我们假设Kcr途径具有一套独特的调节酶、底物和直接结合物，这决定了它与Kac的不同。因此，我们建议使用酶学，化学生物学和蛋白质组学的方法来表征Kcr途径。由于我们的专业知识和我们在过去几年中进行的大量相关研究，我们的团队完全有能力开展这一项目。在这个项目中，我们将首先确定和表征Kcr调节酶，巴豆酰转移酶和decrotonylases，使用各种化学生物学和酶学方法。然后，我们将确定底物的Kcr和研究其动态响应Kcr-调节酶和精子发生过程中，通过定量蛋白质组学方法。最后，我们将鉴定和确认组蛋白Kcr肽的直接结合剂。组蛋白Kcr的新酶和结合剂可能为组蛋白生物学及其在疾病中的作用开辟完全不可预见的富有成效的途径。因此，拟议中的研究应该通过揭示新的表观遗传机制来加速当前染色质生物学的研究，这可能类似于发现组蛋白赖氨酸乙酰化和组蛋白赖氨酸甲基化的Kac和Kme调节酶。