Investigating chromatin control by an Adenovirus histone-like protein

研究腺病毒组蛋白样蛋白对染色质的控制

• 批准号: 8908523
• 负责人: Daphne Christina Avgousti
• 金额: $ 5.42万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8908523
• 关键词: Adenovirus Protein; Adenoviruses; Affect; Antiviral Agents; Appearance; Binding; Binding Sites; Biological Assay; Cell Line; Cell physiology; Cells; ChIP-seq; Child; Chromatin; Chromatin Structure; DNA; DNA Binding; DNA Damage; DNA Repair Pathway; DNA biosynthesis; Digestion; Down-Regulation; Electrophoretic Mobility Shift Assay; Epigenetic Process; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genome; Genomic DNA; Genomics; Goals; Growth; Histones; Host Defense; Human; In Vitro; Infection; Influenza; Investigation; Knowledge; Laboratories; Mass Spectrum Analysis; Micrococcal Nuclease; Mutate; N-terminal; Nuclear Protein; Nucleosomes; Outcome; Outcomes Research; PTPN11 gene; Pathway interactions; Play; Post-Translational Modification Site; Post-Translational Protein Processing; Production; Proteins; Recombinant Proteins; Research; Resources; Role; Series; Signal Pathway; Signal Transduction; Site; Structure; Tail; Testing; Time; Upper Respiratory Infections; Viral; Viral Proteins; Virion; Virus; Virus Diseases; Yeasts; base; biophysical techniques; ds-DNA; insight; irradiation; mutant; novel; obligate intracellular parasite; protein function; public health relevance; reconstitution; research study; response; transcriptome sequencing; viral DNA

 DESCRIPTION (provided by applicant): Viruses are obligate intracellular parasites that rely on the ability to mimic the structure and function of cellular proteins to redirect the cell towards vral production. In this proposal, I focus on a viral protein from Adenovirus called protein VII whose sequence is highly similar to cellular histone proteins. I propose that this viral histone-like protein facilitates viral growth by mimicking histones and manipulating cellular chromatin. When protein VII was first described as histone-like based on primary sequence in the late 1970s, there was little understanding of the role of histone proteins. Since then, in the field of epigenetics has led to an increased appreciation for conserved site-specific post-translational modifications (PTMs) on histone proteins and their impact on cellular processes. The double-stranded DNA genome of Adenovirus is packaged in the virion with histone-like protein VII. My preliminary results show that nuclear protein VII is sufficient to alter cellular chromatin and can down-regulate the DNA damage response (DDR) upon irradiation. I also find that protein VII is modified analogously to histone PTMs, which may be involved in the mechanism of chromatin change or DDR down-regulation. Protein VII PTMs may also serve to disrupt cellular histone PTMs and alter transcription of antiviral genes to promote viral replication. The objective of this proposal is to determine functions of protein VII at cellular chromatin and how these benefit viral replication. I propose to utilize current knowledge of histones as a framework to investigate protein VII function through three specific aims: 1) to define the impact of protein VII on cellula chromatin structure, 2) to define the impact of protein VII on the DNA damage response, and 3) to determine the effect of protein VII on transcription. The outcome of this research will provide a basis for defining the chromatin changes that take place during viral infection. This is the firs time that a histone-like protein has been found to alter cellular chromatin or effect an essential cellular pathway. This project will be a unique contribution to the field of epigenetics as it is te first investigation into the function of a viral histone-like protein in the context of endogenous cellular histone PTMs and the cellular DDR.

 描述(申请人提供)：病毒是专性的细胞内寄生虫，它依赖于模仿细胞蛋白质的结构和功能的能力来将细胞重定向到VAL生产。在这项提议中，我将重点放在来自腺病毒的一种病毒蛋白，称为蛋白VII，其序列与细胞组蛋白高度相似。我认为这种病毒样组蛋白通过模仿组蛋白和操纵细胞染色质来促进病毒的生长。20世纪70年代末，当蛋白质VII首次被描述为基于初级序列的组蛋白样时，人们对组蛋白的作用知之甚少。从那时起，在表观遗传学领域，人们越来越多地认识到组蛋白上保守的位点特异性翻译后修饰(PTM)及其对细胞过程的影响。腺病毒的双链DNA基因组被包装在病毒粒子中，并带有组蛋白样蛋白VII。我的初步结果表明，核蛋白VII足以改变细胞染色质，并可以 下调辐射后DNA损伤反应(DDR)。我还发现，蛋白VII与组蛋白PTM类似地被修饰，这可能参与了染色质改变或DDR下调的机制。蛋白VII PTM也可能破坏细胞组蛋白PTM，改变抗病毒基因的转录以促进病毒复制。这样做的目的是 建议确定细胞染色质中蛋白VII的功能以及这些功能对病毒有何益处。 复制。我建议利用现有的组蛋白知识作为一个框架，通过三个特定的目的来研究蛋白VII的功能：1)确定蛋白VII对细胞染色质结构的影响，2)确定蛋白VII对DNA损伤反应的影响，以及3)确定蛋白VII对转录的影响。这项研究的结果将为确定病毒感染期间发生的染色质变化提供基础。这是第一次发现组蛋白样蛋白可以改变细胞染色质或影响重要的细胞途径。这个项目将是对表观遗传学领域的独特贡献，因为它是第一次在内源性细胞组蛋白PTMS和细胞DDR的背景下研究病毒组蛋白样蛋白的功能。