KSHV latent infection replication

KSHV潜伏感染复制

• 批准号: 9215662
• 负责人: Kenneth M Kaye
• 金额: $ 44.38万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9215662
• 关键词: Acquired Immunodeficiency Syndrome; Africa South of the Sahara; Architecture; Binding; Binding Sites; Biology; Blood group antigen S; Cell Cycle; Cell Nucleus; Cell Survival; Cells; Closure by clamp; Complex; Crystallization; DNA; DNA biosynthesis; DNA-Directed DNA Polymerase; Daughter; Development; Dominant-Negative Mutation; Epidemic; Episome; Epstein-Barr Virus Infections; Etiology; Genome; HIV; Herpesviridae; Herpesviridae Infections; Human; Human Herpesvirus 4; Human Herpesvirus 8; Immune; Kaposi Sarcoma; Latent virus infection phase; Link; Malignant Neoplasms; Mediating; Multicentric Angiofollicular Lymphoid Hyperplasia; Oral cavity; Organ; Outcome; Persons; Plasmids; Polymerase; Process; Proliferating; Proteins; Reagent; Recruitment Activity; Resolution; Roentgen Rays; Role; Saliva; Site; Structure; Terminal Repeat Sequences; Testing; Therapeutic; Validation; Viral; Viral Genome; Virus; Virus Diseases; Virus Latency; Virus Replication; Visceral; Work; antigen binding; cell type; co-infection; daughter cell; experimental study; inhibitor/antagonist; latency-associated nuclear antigen; latent infection; molecular imaging; neoplastic cell; prevent; primary effusion lymphoma; public health relevance; segregation; single molecule; small molecule inhibitor; targeted treatment; therapeutic target; tumor; viral DNA

 DESCRIPTION (provided by applicant): Kaposi's sarcoma (KS)-associated herpesvirus (KSHV or HHV8) is the causative agent of KS, primary effusion lymphoma (PEL) and multicentric Castleman's disease. KSHV is spread person to person through saliva. KSHV tumors occur primarily in AIDS and other immune compromised states. There are no available specific therapies for these tumors. KS is the leading AIDS malignancy and is epidemic in sub Saharan Africa where coinfection with HIV is common. KS often involves the oral cavity and visceral organs. KSHV latently infects tumor cells. During latent infection viral genomes persist as multiple copy, extrachromosomal, circular episomes (plasmids). To persist in proliferating cells, episomes must replicate with each cell cycle and efficiently segregate to daughter nuclei. The latency-associated nuclear antigen (LANA) mediates KSHV episome persistence and is strictly required for viral latency and KSHV survival in proliferating cells. Episome maintanence is comprised of two components: replication of KSHV DNA, and segregation of replicated episomes to daughter cell nuclei. LANA is responsible for both these functions. To replicate KSHV DNA, LANA binds viral terminal repeat (TR) DNA. Since no KSHV DNA replication proteins are expressed during latent infection, LANA is responsible for recruiting host cell replication machinery. Although LANA interacts with several cell replication factors, little is known regarding the underlying mechanisms through which LANA mediates replication. We recently found that LANA recruits the DNA polymerase clamp loader to mediate efficient replication and viral persistence. Loss of this recruitment greatly inhibited LANA's ability to mediate DNA replication and resulted in loss of virus infection. These findings suggested that clamp loading is a rate-limiting step in DNA replication that is incompatible with virus survival. LANA enhancement of clamp loading enables KSHV replication, persistence, and survival. The overarching hypothesis of this application is that LANA's recruitment of the clamp loader is a fundamental component of KSHV replication biology that is an ideal target for potential therapeutic inhibition. Experiments will test this hypothesis. Work will assess the mechanism by which LANA enhances clamp loading. Experiments will investigate the architecture of the LANA-clamp loader complex. In addition, experiments will target the LANA-clamp loader complex for inhibition. The expected outcome of this work is a detailed understanding of the mechanism through which LANA acts on the clamp loader and validation of this interaction as a potential therapeutic target for KSHV malignancy.

 描述（由适用提供）：Kaposi的肉瘤（KS）相关疱疹病毒（KSHV或HHV8）是KS的真正药物，原发性废水淋巴瘤（PEL）和多中心Castleman病。 KSHV是通过唾液传播到个人的。 KSHV肿瘤主要发生在艾滋病和其他免疫受损状态。这些肿瘤没有可用的特定疗法。 KS是领先的艾滋病恶性肿瘤，在撒哈拉以南非洲的流行病很常见。 KS通常涉及口腔和内脏器官。 KSHV潜在感染的肿瘤细胞。在潜在感染期间，病毒基因组持续为多个拷贝，外染色体，循环（质粒）。为了持续存在增殖的细胞，偶发必须在每个细胞周期中复制并有效地分离为核心。潜伏期相关的核抗原（LANA）介导了KSHV发作的持久性，并且严格要求病毒潜伏期和增殖细胞中的KSHV存活。沿着围s的维护由两个组成部分组成：KSHV DNA的复制以及将复制的偶发分离为子细胞核。 Lana负责这两个功能。为了复制KSHV DNA，LANA结合病毒末端重复（TR）DNA。由于在潜在感染期间没有发出KSHV DNA复制蛋白，因此LANA负责募集宿主细胞复制机制。尽管Lana与几个细胞复制因子相互作用，但对于LANA介导复制的基本机制知之甚少。我们最近发现，LANA募集了DNA聚合酶夹具加载器以介导有效的复制和病毒持久性。失去这种招募极大地抑制了Lana介导DNA复制的能力，并导致病毒感染丧失。这些发现表明，夹紧载荷是DNA复制的限制步骤，与病毒存活不符。 LANA增强夹具载荷可以使KSHV复制，持久性和存活率。该应用的总体假设是Lana招募夹具装载机是KSHV复制生物学的基本组成部分，它是潜在的治疗抑制的理想目标。实验将检验这一假设。工作将评估LANA增强夹具载荷的机制。实验将研究LANA-CLAMP装载机复合物的结构。此外，实验将针对LANA-CLAMP装载机复合物进行抑制。这项工作的预期结果是对LANA对夹具装载机作用的机制的详细理解，并将这种相互作用验证为KSHV恶性肿瘤的潜在治疗靶点。