Diversifying the histone code: defining the role of non-canonical phosphorylation in chromatin structure

组蛋白密码多样化：定义非规范磷酸化在染色质结构中的作用

• 批准号: 2749911
• 金额: --
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2749911
• 关键词: Diversifying; histone; code; defining; role

Post-translational modifications (PTMs) of histones are known to direct chromatin structure and thus gene transcription. This so called "histone code" hypothesises that the combination of PTMs on different histones are a driving force that regulate DNA packing and thus the ability of different genes to be accessible for transcription or 'silenced'. Using technology recently developed in our lab for the cell-wide identification of sites of non-canonical phosphorylation (NCP) on e.g. histidine, lysine, arginine using mass spectrometry, we have identified NCP sites on histone proteins (e.g. H1.4, H2B, H3), and on enzymes that themselves regulate histone PTM status (e.g. HDAC1/2/7, CBX1, KAT6A). Identification of these NCP sites raises the possibility that the histone code, as we understand it, may very well be incomplete. The current methods for understanding histone modifications rely on acid-based histone extraction that is incompatible with these types of labile NCP events. Alternatively, PTM site specific antibodies are employed, that due either to epitope incompatibility arising from closely localised PTMs, or different PTMs on the same site, may bias enrichment and preclude identification. This project aims to re-define the histone code and its regulation, using methods that are compatible with retaining sites of acid-labile phosphorylation on histones and their regulatory enzymes. We will also explore the relationship between this re-defined histone code and chromatin structure.

组蛋白的翻译后修饰（PTMs）被认为是直接染色质结构和基因转录。这种所谓的“组蛋白密码”假设，不同组蛋白上ptm的组合是调节DNA包装的驱动力，从而使不同基因能够被转录或“沉默”。利用我们实验室最近开发的技术，利用质谱技术在细胞范围内鉴定非规范磷酸化（NCP）位点，如组氨酸、赖氨酸、精氨酸，我们已经鉴定了组蛋白上的NCP位点（如H1.4、H2B、H3），以及自身调节组蛋白PTM状态的酶上的NCP位点（如HDAC1/2/7、CBX1、KAT6A）。这些新冠病毒位点的鉴定提出了一种可能性，即我们所理解的组蛋白代码很可能是不完整的。目前了解组蛋白修饰的方法依赖于酸基组蛋白提取，这与这些不稳定的NCP事件不相容。或者，使用PTM位点特异性抗体，由于紧密定位的PTMs引起的表位不相容，或同一位点上的不同PTMs，可能会偏向富集并排除鉴定。该项目旨在重新定义组蛋白编码及其调控，使用与保留组蛋白及其调控酶上酸不稳定磷酸化位点相容的方法。我们还将探讨这种重新定义的组蛋白密码和染色质结构之间的关系。