Modulation of histone deacetylase activity by a gammaherpesviral kinase

伽马疱疹病毒激酶对组蛋白脱乙酰酶活性的调节

• 批准号: 8524237
• 负责人: Bryan C Mounce
• 金额: $ 3.31万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2014-03-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8524237
• 关键词: Address; Alleles; Antiviral Agents; Biological Assay; Biological Models; Cell Nucleus; Cells; Chromatin; Clinical Trials; DNA; DNA Viruses; DNA biosynthesis; Data; Development; Disease; Drug resistance; Enzymes; Estrogen Receptors; Excision; Exclusion; Family; Fibroblasts; Frequencies; Gene Expression; Genetic; Genetic Transcription; Genome; HDAC1 gene; Herpesviridae; Herpesvirus 1; Histone Acetylation; Histone Deacetylase; Histones; Human; Human Herpesvirus 4; Human Herpesvirus 8; Human Virus; In Vitro; Infection; Integration Host Factors; Kaposi Sarcoma; Life; Lung; Lymphoproliferative Disorders; Malignant Neoplasms; Measures; Mediating; Modeling; Monitor; Mus; Names; Nasopharynx Carcinoma; Nature; Nuclear; Pathogenesis; Phosphotransferases; Physiological; Plaque Assay; Play; Post-Translational Protein Processing; Production; Protein Isoforms; Protein Kinase; Proteins; Publishing; RNA Interference; Role; Spleen; Staging; Substrate Interaction; System; Techniques; Testing; Therapeutic; Translating; Viral; Viral Genes; Viral Genome; Viral Pathogenesis; Viral Proteins; Virion; Virus; Virus Diseases; Virus Latency; Work; base; cancer therapy; gammaherpesvirus; in vivo; inhibitor/antagonist; latent infection; monolayer; pathogen; promoter; public health relevance; reactivation from latency; recombinase; success; therapeutic development; therapeutic target

DESCRIPTION (provided by applicant): Gammaherpesviruses are pervasive pathogens that establish lifelong infection in their hosts. The human gammaherpesviruses Epstein-Barr Virus (EBV) and Kaposi's Sarcoma Associated Herpesvirus (KSHV) are associated with diverse diseases, including several cancers. The murine gammaherpesvirus named mouse gammaherpesvirus 68 (MHV68) serves as a model system and provides for the use of powerful mouse genetics to understand the virus-host relationship. All gammaherpesviruses encode a viral protein kinase that phosphorylates and interacts with a multitude of substrates, though the physiological relevance of many of these substrate interactions remains to be verified. As nuclear DNA viruses, herpesviruses interact with cellular histone proteins that enwrap DNA in the nucleus to form chromatin. Histone acetylation is a common post-translational modification that promotes enhanced gene expression. Histone deacetylases (HDACs) catalyze the removal of acetyl moieties from histones and are classically considered to repress transcription. Due to their repressive nature, HDACs are frequently characterized as antiviral, and several viruses have evolved mechanisms to inactivate, disrupt, relocalize, or modify HDACs to facilitate infection. For example, herpes simplex virus-1 (HSV-1) has evolved mechanisms to antagonize HDACs through two distinct viral proteins: ICP02 and the Us3 kinase3;4. We have discovered that orf36, the MHV68 kinase, interacts with and modulates HDACs during infection. We hypothesize that this modulation facilitates viral infection at several stages, including viral gen expression, DNA replication, and infectious virion production. To test this hypothesis, we will use cells and mice with conditional HDAC alleles. After excision of HDACs in cells in vitro, we will measure viral gene expression and DNA replication via qRT-PCR and infectious virion production via plaque assay. After HDAC excision in vivo, we will measure peak viral titers via plaque assay and parameters of viral latency with limiting dilution assays. The combination of in vitro and in vivo techniques will encompass a directed, yet extensive, characterization of the role of HDACs in MHV68 infection. Herpesviral kinases are critical regulators of infection and represent a tractable target for therapeutics. Additionally, histone deacetylases have emerged as important therapeutic targets in cancer treatments. Thus, understanding the interactions between herpesviral kinases, chromatin, and chromatin-modifying enzymes is critical to the development of rational therapeutics. Several orf36 functions are conserved in other gammaherpesviral kinases; thus, the results of our studies are expected to translate to human pathogens.

描述（由申请方提供）：γ疱疹病毒是在其宿主中建立终身感染的普遍病原体。人类γ疱疹病毒Epstein-Barr病毒（EBV）和卡波西肉瘤相关疱疹病毒（KSHV）与多种疾病相关，包括几种癌症。小鼠γ疱疹病毒命名为小鼠γ疱疹病毒68（MHV 68）作为一个模型系统，并提供了强大的小鼠遗传学的使用，以了解病毒-宿主的关系。所有的γ疱疹病毒编码的病毒蛋白激酶磷酸化和与多种底物相互作用，虽然许多这些底物相互作用的生理相关性仍有待验证。作为核DNA病毒，疱疹病毒与将DNA包裹在细胞核中的细胞组蛋白相互作用以形成染色质。组蛋白乙酰化是一种常见的翻译后修饰，可促进基因表达增强。组蛋白脱乙酰酶（HDAC）催化乙酰基部分从组蛋白的去除，并且经典地被认为抑制转录。由于它们的抑制性质，HDAC通常被表征为抗病毒的，并且几种病毒已经进化出破坏、破坏、重新定位或修饰HDAC以促进感染的机制。例如，单纯疱疹病毒-1（HSV-1）已经进化出通过两种不同的病毒蛋白质拮抗HDAC的机制：ICP 02和Us 3激酶3;4。我们已经发现，orf 36，MHV 68激酶，在感染过程中与HDAC相互作用并调节HDAC。我们假设这种调节在几个阶段促进了病毒感染，包括病毒基因表达、DNA复制和感染性病毒体产生。为了验证这一假设，我们将使用具有条件性HDAC等位基因的细胞和小鼠。在体外切除细胞中的HDAC后，我们将通过qRT-PCR测量病毒基因表达和DNA复制，并通过空斑测定测量感染性病毒体产生。在体内HDAC切除后，我们将通过空斑测定法测量峰值病毒滴度，并通过有限稀释测定法测量病毒潜伏期参数。体外和体内技术的结合将包括一个有针对性的，但广泛的，表征的作用， HDAC在MHV 68感染中的作用。疱疹病毒激酶是感染的关键调节因子，并且代表治疗的易处理靶标。此外，组蛋白脱乙酰酶已成为癌症治疗中的重要治疗靶标。因此，了解疱疹病毒激酶，染色质，染色质修饰酶之间的相互作用是至关重要的合理治疗的发展。几个orf 36功能在其他γ疱疹病毒激酶中是保守的，因此，我们的研究结果有望转化为人类病原体。