Studies on Epigenetically Active Latent Chromatin Maintenance

表观遗传活性潜在染色质维持的研究

基本信息

批准号：
10570202
负责人：
Yoshihiro Izumiya
金额：
$ 52.98万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-10 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10570202
关键词：
Affinity Chromatography Alanine Alkylation Amino Acid Sequence Authorization documentation Bacterial Artificial Chromosomes Baculoviruses Binding Biochemical Biological Phenomena CHD4 gene Cell Line Cell Lineage Cells Chromatin Chromatin Structure Clinical Complex Cryoelectron Microscopy DNA DNA biosynthesis Disease Docking Engineering Enhancers Enzymes Epigenetic Process Episome Event Frequencies Gene Activation Genes Genetic Genetic Enhancer Element Genetic Transcription Genetic study Genome Genomic Segment Genomics Herpesviridae Hi-C Human Herpesvirus 4 Human Herpesvirus 8 Infection Laboratories Life Cycle Stages Link Lymphoma Lytic Maintenance Malignant Neoplasms Mammalian Cell Maps Mediating Memory Metabolic Modeling Molecular Mutation Nature Noise Nuclear Organism Outcome Peptides Pharmaceutical Preparations Phenotype Play Population Proteins Proteomics RNA RNA Polymerase II Recombinant DNA Recombinants Regulation Regulatory Element Resolution Risk Factors Role Signal Transduction Site Small RNA Stimulus Structure Sulfhydryl Compounds Surface Terminal Repeat Sequences Therapeutic Intervention Tissues Trans-Activators Transcription Initiation Site Untranslated RNA Viral Virus Latency authority biosafety level 2 facility cell killing cellular targeting cost effective disease phenotype ds-DNA infected B cell insight institutional biosafety committee latency-associated nuclear antigen latent infection lytic gene expression lytic replication meter mutant nuclease peptide drug promoter protein complex stem tissue culture tool transcription factor transcriptome sequencing transcriptomics tumor tumorigenesis yeast genetics

项目摘要

Deregulation of genome interactions through mutations at regulatory elements or mutation of responsible cellular nuclear enzymes result in multiple disorders. Although the significance of proper genomic-genomic interactions has been seen from disease phenotypes, the underlying molecular mechanisms of genomic looping regulation and the direct outcome of transcription are still not well connected. This partly stems from the highly complex nature of cellular promoter regulation, since it is controlled by multiple transcriptional factors and enhancer elements with significant noise within cell populations. Transcriptomics, genomics, and proteomics studies with recombinant Kaposi's sarcoma-associated herpesvirus (KSHV) episomes identified that the host cellular ChAHP (CHD4, ADNP, HP1) protein complex forms a stable protein complex with KSHV latency associated nuclear antigen (LANA) and plays essential roles in maintaining the inducibility of latent viral chromatin. The ChAHP complex is known to restrict cellular enhancer accessibility and regulates cell lineage, although how the protein complex regulates enhancer-promoter interaction remains unknown. Here, we hypothesize that the KSHV latency-lytic switch is a product of enhancer-promoter interactions regulated by the LANA-ChAHP complex. In this application, we will study how the KSHV episome is maintained as an inducible episome and if we can target the protein complex for therapeutic intervention. The molecular action of the LANA/ChAHP protein complex will be studied from three different perspectives; (i) biochemical, (ii) genetic, and (iii) protein complex structure. By taking advantage of defined enhancer-promoter pairs and convenient inducible mini viral chromatin, we will study enhancer-promoter regulations by dissecting ChAHP protein complex function. The proposed studies should not only benefit the understanding of the KSHV latency-lytic switch, but also provide insight into cellular inducible enhancer regulatory mechanisms.

通过调控元件的突变或负责基因的突变来解除基因组相互作用的管制细胞核酶会导致多种疾病。尽管正确的基因组学的重要性从疾病表型、基因组循环的潜在分子机制中可以看出相互作用调控和转录的直接结果仍然没有很好地联系起来。这部分源于高度细胞启动子调控的复杂性，因为它受到多种转录因子和增强子的控制细胞群中具有显着噪音的元素。重组卡波西肉瘤相关的转录组学、基因组学和蛋白质组学研究疱疹病毒 (KSHV) 附加体鉴定出宿主细胞 ChAHP (CHD4、ADNP、HP1) 蛋白复合物形成与 KSHV 潜伏期相关核抗原 (LANA) 的稳定蛋白复合物，在维持潜伏病毒染色质的诱导能力。 ChAHP 复合体已知会限制细胞增强子的可及性调节细胞谱系，尽管蛋白质复合物如何调节增强子-启动子相互作用仍然未知。在这里，我们假设 KSHV 潜伏裂解开关是增强子-启动子相互作用的产物，受以下因素调节： LANA-ChAHP 复合体。在此应用中，我们将研究 KSHV 附加体如何维持为诱导型附加体以及我们是否可以针对蛋白质复合物进行治疗干预。的分子作用 LANA/ChAHP蛋白复合物将从三个不同的角度进行研究； (i) 生化，(ii) 遗传，以及 (iii) 蛋白质复合物结构。通过利用确定的增强子-启动子对和方便的诱导型迷你病毒染色质，我们将通过剖析 ChAHP 蛋白复合物功能来研究增强子-启动子调控。这拟议的研究不仅应该有助于理解 KSHV 潜伏裂解开关，而且还可以提供见解进入细胞诱导增强子调节机制。