Studying chromosome function using chemical biology

利用化学生物学研究染色体功能

• 批准号: 8161780
• 负责人: TARUN M. KAPOOR
• 金额: $ 41.47万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8161780
• 关键词: Acetylation; Alkynes; Benzophenones; Binding; Biochemical; Biochemistry; Biological; Biology; Cell division; Cell physiology; Cells; Cellular biology; Chemicals; Chemistry; Chromatin; Chromosome Segregation; Chromosomes; Code; Complex; DNA; DNA Sequence; DNA biosynthesis; Data; Disease; Ensure; Epigenetic Process; Gene Expression; Genetic Transcription; Genome; Goals; Histone Code; Histone H3; Histones; Human; Knowledge; Lead; Link; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Methodology; Methods; Methylation; Microscopy; Mitosis; Modification; Names; Normal Cell; Peptides; Pharmaceutical Preparations; Phosphorylation; Play; Post-Translational Protein Processing; Process; Protein Binding; Proteins; Proteomics; Publishing; Reader; Reading; Recruitment Activity; Reporting; Research; Research Proposals; Resolution; Site; Stream; Tail; Testing; Therapeutic; Work; Writing; aurora kinase; base; cancer cell; cell type; chromatin protein; crosslink; design; histone modification; improved; insight; novel therapeutics; protein function; protein profiling; protein protein interaction; repaired; segregation; trait; transmission process; ubiquitin ligase

DESCRIPTION (provided by applicant): The fact that DNA is wrapped around a spool, comprised of histones, is believed to influence essentially all aspects of chromosome biology, including DNA replication, repair after damage and segregation. Diverse post-translational modifications (e.g. acetylation, methylation, and phosphorylation) of histones are known and are believed to play key roles in regulating a wide swath of biology linked to our genomes. Based on observed antagonisms and synergies between different histone post-translational modifications (or 'marks') in recruiting proteins to chromosomes, it has been proposed that these 'marks' form a 'code' for regulating chromosome function. It has also been suggested that this 'code' may provide a basis of epigenetic inheritance, which is the transmission of cellular traits that are not encoded at the level of DNA sequence. Many of the proteins that post-translationally modify histones (i.e. 'write' or 'erase' the 'code') have been characterized. In contrast, our knowledge of the proteins that recognize (or 'read') histone post-translational modifications remains incomplete. The difficulty in identifying these effector-proteins (or 'readers') is, in large part, due to the histone modifications being sub-stoichiometric, dynamic, and mediators of weak interactions. With the goal to fill this knowledge gap, we have recently reported an approach, which combines photo-chemical crosslinking with bio-orthogonal chemistry, to 'capture' proteins that bind histone H3 trimethylated at Lys-4. We now combine this method with state-of-the-art mass spectrometry to develop a robust chemical proteomics approach to profile 'readers' of histone methylation 'marks.' Our ongoing work suggests that our approach is general and can be used to analyze these post-translational modification-dependent protein-protein interactions in any human cell type (e.g. normal or cancer), cell state (e.g. mitosis) or context (e.g. drug-treated). Based on these and other unpublished preliminary data, we propose to: (i) comprehensively profile proteins that recognize methylation 'marks' on histones, (ii) characterize how proteins that recognize methylation 'marks' control down-stream biology, and (iii) examine how interplay between histone phosphorylation and methylation ensures error-free chromosome segregation during cell division. We combine chemistry, biochemistry, high-resolution microscopy and cell biological approaches to gain insight into fundamental cellular processes. Our findings may reveal how improper 'reading' of histone post-translational modifications can result in disease. In the long-term, our findings may also provide a basis for developing new therapeutic strategies that target 'readers' of histone methylation 'marks'. PUBLIC HEALTH RELEVANCE: The goals of the proposed research are to develop and apply a chemical proteomics approach to comprehensively profile proteins that recognize histones with particular post-translational modifications (or 'marks'). By characterizing these proteins, we will gain insight into how chromosome function is regulated in normal cells and how diseases may arise when the 'reading' of these histone 'marks' goes awry.

描述（由申请人提供）：据信，将DNA包裹在阀芯周围的事实，该事实被认为会影响染色体生物学的所有方面，包括DNA复制，损伤后修复和隔离。已知组蛋白的各种翻译后修饰（例如乙酰化，甲基化和磷酸化）已知，并且被认为在调节与我们的基因组相关的广泛生物学方面起着关键作用。基于观察到的拮抗作用和在募集蛋白质到染色体的不同组蛋白后翻译后修饰（或“标记”）之间的拮抗作用和协同作用，已经提出，这些“标记”形成了调节染色体功能的“代码”。还有人建议该“代码”可以提供表观遗传遗传的基础，这是在DNA序列水平上未编码的细胞性状的传播。经过翻译后修改组蛋白的许多蛋白质（即“写”或“擦除”“代码”）的表征已经表征。相反，我们对识别（或“读取”）组蛋白的蛋白质的了解仍然不完整。在很大程度上，由于组蛋白的修饰是弱相互作用的亚化学计量，动态和介体，因此很难识别这些效应蛋白（或“读者”）。为了填补这一知识差距，我们最近报告了一种方法，该方法将光化交联与生物正交化学结合在一起，以“捕获”在Lys-4处结合组蛋白H3三甲基化的蛋白质。现在，我们将此方法与最先进的质谱法相结合，以开发出强大的化学蛋白质组学方法来介绍组蛋白甲基化的“读者”“标记”。我们正在进行的工作表明我们的方法是一般的，可用于分析任何人类细胞类型（例如正常或癌症），细胞状态（例如有丝分裂）或上下文（例如药物治疗）中这些翻译后修饰依赖性蛋白 - 蛋白质相互作用。 Based on these and other unpublished preliminary data, we propose to: (i) comprehensively profile proteins that recognize methylation 'marks' on histones, (ii) characterize how proteins that recognize methylation 'marks' control down-stream biology, and (iii) examine how interplay between histone phosphorylation and methylation ensures error-free chromosome segregation during cell division.我们结合了化学，生物化学，高分辨率显微镜和细胞生物学方法，以深入了解基本的细胞过程。我们的发现可能揭示了组蛋白后翻译后修饰的“阅读”如何导致疾病。从长远来看，我们的发现还可以为开发针对组蛋白甲基化读者“标记”的新的治疗策略提供基础。 公共卫生相关性：拟议研究的目标是开发和应用化学蛋白质组学方法来全面介绍蛋白质，这些蛋白质识别具有特定翻译后修饰的组蛋白（或“标记”）。通过表征这些蛋白质，我们将深入了解在正常细胞中如何调节染色体功能，以及当这些组蛋白“标记”的“读数”出现时，如何出现疾病。