Functional consequences of evolutionary innovation in histone repertoires

组蛋白库进化创新的功能后果

• 批准号: 10644921
• 负责人: Pravrutha Raman
• 金额: $ 12.5万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644921
• 关键词: Affect; Biological; Biological Assay; Biological Process; Biology; Birth; Cell physiology; Cells; Chromatin; DNA; DNA Damage; DNA Packaging; DNA Repair; DNA Repair Gene; Defect; Development; Disease; Drosophila genome; Drosophila genus; Engineering; Eukaryota; Event; Evolution; Fertility; Foundations; Frequencies; Future; Gene Duplication; Gene Expression; Gene Expression Regulation; Gene Fusion; Genes; Genetic; Genetic Transcription; Genome; Genomics; Growth; Heterochromatin; Histone Fold; Histone H2A; Histones; Learning; Length; Link; Location; Malignant Neoplasms; Mediating; Meiosis; Mitosis; Molecular Evolution; Mutate; Mutation; N-terminal; Organism; Partner in relationship; Phenotype; Phylogenetic Analysis; Play; Process; Protein Family; Proteins; Recording of previous events; Recurrence; Regulation; Research; Role; Saccharomyces cerevisiae; Saccharomycetales; Specificity; Tail; Testing; Training; Variant; Work; Yeasts; fitness; flexibility; fly; fungus; gene repression; genomic locus; improved; in vivo; innovation; insight; model organism; mutant; novel; pressure; prevent; programs; response; skills; tool; Affect; Biological; Biological Assay; Biological Process; Biology; Birth; Cell physiology; Cells; Chromatin; DNA; DNA Damage; DNA Packaging; DNA Repair; DNA Repair Gene; Defect; Development; Disease; Drosophila genome; Drosophila genus; Engineering; Eukaryota; Event; Evolution; Fertility; Foundations; Frequencies; Future; Gene Duplication; Gene Expression; Gene Expression Regulation; Gene Fusion; Genes; Genetic; Genetic Transcription; Genome; Genomics; Growth; Heterochromatin; Histone Fold; Histone H2A; Histones; Learning; Length; Link; Location; Malignant Neoplasms; Mediating; Meiosis; Mitosis; Molecular Evolution; Mutate; Mutation; N-terminal; Organism; Partner in relationship; Phenotype; Phylogenetic Analysis; Play; Process; Protein Family; Proteins; Recording of previous events; Recurrence; Regulation; Research; Role; Saccharomyces cerevisiae; Saccharomycetales; Specificity; Tail; Testing; Training; Variant; Work; Yeasts; fitness; flexibility; fly; fungus; gene repression; genomic locus; improved; in vivo; innovation; insight; model organism; mutant; novel; pressure; prevent; programs; response; skills; tool

PROJECT SUMMARY Histone proteins package DNA into chromatin and regulate all DNA-templated biological processes in eukaryotes. Consistent with their essential function, mutations or misregulation of histones result in many diseases. While core histones primarily function in genome packaging, histone variants can replace canonical histones at unique genomic locations for specialized roles such as DNA damage response (DDR) or gene expression. Despite their essential functions, histone repertoires have undergone distinct lineage-specific changes. Such evolutionary novelty via gene fusions, duplications or sequence divergence is unexpected in conserved protein families and suggestive of an adaptive advantage for sequence innovation. This proposal will use evolutionary innovations in eukaryotic histone H2A repertoires to investigate the causes and consequences of evolutionary turnover of histone proteins. The most common eukaryotic H2A repertoire is made up of core histone H2A, and histone variants H2A.X and H2A.Z that are involved in DDR and gene regulation, respectively. However, two evolutionary shifts, both likely selectively advantageous, have occurred in yeast and Drosophila H2A repertoires. In yeast, H2A.X, entirely replaced canonical H2A, likely improving DDR and affecting processes like meiosis that depend on DDR. In Drosophila, H2A.X fused with H2A.Z giving rise to a unique H2Av variant. This fusion enriches DDR at heterochromatin which potentially restricts DDR- based transposition events to gene poor regions. To identify functional consequences of these evolutionary innovations, the applicant will re-engineer the ancestral eukaryotic H2A repertoire in yeast and flies. Specifically in S. cerevisiae, the applicant will engineer a core H2A and two variants H2A.X and H2A.Z, preventing H2A.X from being the core histone (Aim 1). In D. melanogaster, the fusion histone H2Av will be separated into H2A.X and H2A.Z uncoupling DDR and gene regulation functions (Aim 2). Changes to organismal chromatin packaging, and relevant biological functions including DNA repair, meiosis, fertility, and transposition will be interrogated. In Aim 3, the applicant will apply tools learnt in Aims 1 and 2 to study divergence of eukaryotic core H2A. Histones have tail sequences which are heavily post-translationally modified and play crucial roles for higher order chromatin packaging and protein interactions. The high sequence divergence across eukaryotic histone tails suggests that tails could facilitate unique lineage-specific functions. By engineering different tail sequences in yeast and flies, Aim 3 will reveal changes to chromatin packaging, and changes to processes such as mating, and quiescence in yeast, and fertility and development in flies. To launch this work, the applicant requires training in genetics, genomics, and phenotypic assays in two model organisms, yeast and flies. By leveraging, her expertise in evolutionary analyses and the power of well-established tools in yeast and flies simultaneously, in the future the applicant will study the biological basis and consequences of histone innovation including her own previous discoveries and co-evolving mechanisms.

项目摘要 组蛋白蛋白将DNA包装到染色质中，并调节所有DNA模拟的生物学过程 真核生物。与它们的基本功能，突变或组蛋白的不调节一致，导致许多 疾病。虽然核心组蛋白主要在基因组包装中起作用，但组蛋白变体可以替代规范 在独特的基因组位置的组蛋白，用于专业角色，例如DNA损伤反应（DDR）或基因 表达。尽管它们具有重要的功能，但组蛋白曲目还是经历了明显的谱系特异性 更改。这种进化的新颖性通过基因融合，重复或序列发散是出乎意料的 保守的蛋白质家族，并提出了序列创新的适应性优势。这个建议 将在真核组蛋白H2A曲目中使用进化创新来研究原因和 组蛋白进化周转的后果。最常见的真核H2A曲目是 由DDR和基因涉及的核心组蛋白H2A和组蛋白变体H2A.X和H2A.Z组成 调节。但是，两次进化变化都可能有选择性地有利， 在酵母和果蝇H2A曲目中。在酵母中，H2A.X，完全替换了规范H2A，可能会改善 DDR和影响依赖DDR的减数分裂的过程。在果蝇中，H2A.X与H2A.Z融合 升至独特的H2AV变体。这种融合富集了异染色质的DDR，这可能限制了DDR- 基于基因较差区域的基于转置事件。确定这些进化的功能后果 创新，申请人将在酵母和苍蝇中重新设计祖先真核H2A曲目。 特别是在酿酒酵母中，申请人将设计一个核心H2A和两个变体H2A.X和H2A.Z， 防止H2A.X成为核心组蛋白（AIM 1）。在D. Melanogaster中，融合组蛋白H2AV将是 分为H2A.X和H2A.Z解耦合DDR和基因调节函数（AIM 2）。更改为 有机染色质包装以及相关的生物学功能，包括DNA修复，减数分裂，生育和 转座将受到审问。在AIM 3中，申请人将应用目标1和2中学到的工具来学习 真核核心H2A的发散。组蛋白的尾部序列在后翻译上很大 修改并扮演高阶染色质包装和蛋白质相互作用的关键作用。高 真核组蛋白尾巴的序列差异表明，尾巴可以促进独特的谱系特异性 功能。通过在酵母和果蝇中工程进行不同的尾序，AIM 3将揭示染色质的变化 包装，以及诸如酵母交配和静止的过程以及生育和发育 在苍蝇中。为了启动这项工作，申请人需要在遗传学，基因组学和表型测定中进行培训 两个模型生物，酵母和苍蝇。通过利用她在进化分析方面的专业知识和 酵母中建立良好的工具，并同时飞行，将来申请人将研究生物学基础 组蛋白创新的后果，包括她自己以前的发现和共同发展的机制。