Genetic and Biochemical Studies of KSHV LANA

KSHV LANA 的遗传和生化研究

基本信息

批准号：
10599894
负责人：
Kenneth M Kaye
金额：
$ 65.55万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10599894
关键词：
Acquired Immunodeficiency Syndrome Africa South of the Sahara Animal Model Binding Biochemical Biological Biological Assay C-terminal Cell Nucleus Cell Proliferation Cell Survival Cells DNA DNA Binding Domain DNA biosynthesis Daughter Development Disease Elements Epidemic Episome Etiology Genes Genetic Genome Glycogen Synthase Kinase 3 Herpesviridae Infections Histone H2A Human Human Herpesvirus 8 Immunocompromised Host Investigation Kaposi Sarcoma Link Lysine Maintenance Malignant Neoplasms Mediating Mitotic Chromosome Molecular Multicentric Angiofollicular Lymphoid Hyperplasia Mus N-terminal Nucleosomes Oncoproteins Oral cavity Organ Phosphorylation Site Plasmids Proliferating Proteins Reader Repetitive Sequence Role Sequence Deletion Sequence Homologs Specificity Surface TP53 gene Terminal Repeat Sequences Viral Viral Genome Virus Virus Replication Visceral Work antigen binding daughter cell experimental study gamma-2 herpesvirus gammaherpesvirus genetic approach in vivo insight interest latency-associated nuclear antigen neoplastic cell novel novel strategies prevent primary effusion lymphoma segregation tumor ubiquitin-protein ligase viral DNA

项目摘要

Kaposi's sarcoma (KS)-associated herpesvirus (KSHV or HHV8) is the causative agent of KS, primary effusion lymphoma (PEL) and multicentric Castleman's disease. These tumors occur in AIDS and other immunocompromised states. There are no specific therapies for these diseases. KS is the leading AIDS malignancy, and is epidemic in sub Saharan Africa. KS often involves the oral cavity and can disseminate to visceral organs. KSHV latently infects tumor cells and viral genomes persist as circular, multiple copy, extrachromosomal, episomes (plasmids). In order to persist in proliferating cells, episomes must first replicate and then efficiently segregate to daughter nuclei. Tumor cell viability is dependent on latent KSHV infection. The latency-associated nuclear antigen (LANA) mediates KSHV episome persistence and this function is essential for virus survival. Therefore, LANA function is of fundamental importance. LANA represents the Achilles heel of KSHV as its disruption is lethal to the virus. LANA is necessary and sufficient for KSHV episome maintenance in the absence of other virus genes. Episome persistence is comprised of two components: 1) replication of viral DNA and 2) segregation of the replicated episomes to progeny cell nuclei. LANA mediates both these functions. C-terminal LANA binds specific sequence in KSHV terminal repeat (TR) DNA to mediate DNA replication. LANA tethers TR DNA to mitotic chromosomes to segregate genomes to daughter nuclei. To form a molecular tether, the C-LANA DNA binding domain (DBD) binds TR DNA and N-LANA simultaneously binds histones H2A/H2B at the folded portion of the nucleosome. In addition to N- and C-LANA, internal LANA sequence is also critical for episome persistence. We have discovered two distinct internal LANA regions, one comprised of unique sequence, and one within repetitive sequence, that are critical for LANA mediated episome persistence. The unique sequence contains three distinct functional motifs, whereas the repetitive region is homologous to a host cell “reader” sequence. Both of these regions will be investigated in this work. This work will use a detailed, in depth approach to investigate each of the recently identified LANA functional sequences. Experiments will use a rigorously developed panel of assays to identify the functional motif and determine the mechanism through which the unique sequence mediates episome persistence. The repetitive LANA effector sequence homologous to host “reader” sequence will be investigated to determine the mechanism through which it exerts its function. This work will provide important insight into LANA’s fundamental function of episome maintenance, and may lead to novel strategies to prevent or treat KSHV associated malignancies.

卡波西肉瘤 (KS) 相关疱疹病毒（KSHV 或 HHV8）是 KS 的病原体，原发性渗出性淋巴瘤（PEL）和多中心卡斯尔曼病这些肿瘤发生在艾滋病和其他疾病中。 KS 是主要的艾滋病，目前尚无针对这些疾病的具体治疗方法。 KS 是一种恶性肿瘤，在撒哈拉以南非洲地区流行，通常累及口腔，并可传播到口腔。 KSHV 潜伏感染肿瘤细胞，病毒基因组以环状、多拷贝形式存在。染色体外的附加体（质粒）为了在增殖细胞中持续存在，附加体必须首先进行复制。然后有效分离到子细胞核肿瘤细胞的活力取决于潜伏的 KSHV 感染。潜伏期相关核抗原 (LANA) 介导 KSHV 游离体持久性及其功能因此，LANA功能对于病毒的生存至关重要。 KSHV 的致命弱点是 LANA 的破坏对于 KSHV 来说是必要且充分的。在没有其他病毒基因的情况下维持附加体。附加体持久性由两个部分组成：1) 病毒 DNA 的复制和 2) 病毒 DNA 的分离复制到子代细胞核的附加体介导这两种功能。 KSHV 末端重复 (TR) DNA 中的特定序列介导 DNA 复制。有丝分裂染色体将基因组分离到子核，形成分子系链 C-LANA DNA。结合域 (DBD) 结合 TR DNA，N-LANA 在折叠处同时结合组蛋白 H2A/H2B 除了 N- 和 C-LANA 之外，内部 LANA 序列对于附加体也至关重要。坚持。我们发现了两个不同的内部 LANA 区域，一个由独特的序列组成，另一个由在重复序列中，这对于 LANA 介导的附加体持久性至关重要。包含三个不同的功能基序，而重复区域与宿主细胞“阅读器”同源本工作将研究这两个区域。这项工作将使用详细、深入的方法来调查最近识别的每个 LANA 实验将使用严格开发的检测组来鉴定功能序列。基序并确定独特序列介导附加体持久性的机制。将研究与宿主“阅读器”序列同源的重复 LANA 效应子序列，以确定这项工作将为 LANA 提供重要的见解。附加体维持的基本功能，并可能导致预防或治疗 KSHV 的新策略相关恶性肿瘤。