Lana and Cellular Gene Expression in Kaposis Sarcoma

卡波西肉瘤中的 Lana 和细胞基因表达

• 批准号: 7224919
• 负责人: ROLF F RENNE
• 金额: $ 22.67万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7224919
• 关键词: Address; Affinity Chromatography; Binding; Binding Sites; Biochemical; Biological Assay; Biology; C-terminal; Cell Cycle Arrest; Cell Cycle Regulation; Cell Line; Cells; Chromatin; Chromosomes; Cosmids; DNA Binding; DNA Binding Domain; DNA biosynthesis; Disease; EBNA-1 protein; Elements; Environment; Episome; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Genome; Herpesviridae; Homologous Gene; Human; Human Herpesvirus 4; Human Herpesvirus 8; Lymphoid Cell; Lymphoma; Lymphoproliferative Disorders; Maintenance; Malignant Neoplasms; Maps; Mitotic Chromosome; Mutagenesis; Numbers; Pathway interactions; Plasmids; Play; Precipitation; Process; Protein Binding; Protein p53; Proteins; Proteomics; Replication Origin; Role; Rolfing; Signal Transduction; Site; Specificity; TP53 gene; TR-2 mycotoxin; Terminal Repeat Sequences; Transcriptional Regulation; Translating; Tumor Suppressor Proteins; Viral Genes; Viral Genome; Viral Proteins; Work; antigen binding; chromatin remodeling; effusion; latency-associated nuclear antigen; neoplastic cell; origin recognition complex; replicator; sarcoma; segregation

DESCRIPTION (provided by applicant): Kaposi's sarcoma-associated herpesvirus (KSHV) also called Human herpes virus type 8 (HHV-8) is associated with Kaposis's sarcoma (KS), and two lymphoproliferative diseases: Primary effusion lymphomas (PEL) and a subset of Multicentric Castlemen's disease (MCD). Common to these malignancies is that the majority of tumor cells are latently infected and express only a small number of viral genes. One of these genes, ORF73, encodes the latency-associated nuclear antigen (LANA). LANA is highly expressed in all KS tumor cells and plays an important role in the biology of KSHV. LANA is the only viral protein required for latent DNA replication and genome maintenance during latency. In addition, LANA interacts with a variety of cellular proteins including the tumor suppressors p53 and Rb thereby modulating gene expression in latently infected cells. To analyze LANA's ability to modulate cellular gene expression, we performed gene expression profiling and found that LANA regulates a number of genes in the Rb/E2F pathway which, as a consequence can protect lymphoid cells from p161NK4a-induced cell cycle arrest. To study LANA's role in DNA replication we performed a detailed biochemical analysis of its DNA binding specificity and activity. We identified two binding sites within the terminal repeat of the KSHV genome and demonstrated that both sites contribute to the ability of LANA to support replication of TR-containing plasmids. We hypothesize that LANA and/or cis-regulatory sequences within TR interact with cellular proteins that facilitate DNA replication. To address this hypothesis we propose the following three aims that are focused on deciphering mechanisms by which LANA supports DNA replication: 1) Mapping cis-regulatory sequences of the putative origin of replication within TR 2) Mapping trans-requirements for DNA binding and LANA-dependent DNA replication 3) Identification of cellular proteins that interact with LANA-C and/or a minimal origin of replication.

描述（由申请人提供）：Kaposi的肉瘤相关疱疹病毒（KSHV）也称为人类疱疹病毒8型（HHV-8）与Kaposis的肉瘤（KS）和两种淋巴细胞增生性疾病有关：主要淋巴瘤（PEL）（PEL）和一份多症患者（MCCD）的主要淋巴瘤（PEL）。这些恶性肿瘤共有的是，大多数肿瘤细胞受到潜在感染，并且仅表达少量的病毒基因。这些基因之一ORF73编码与潜伏期相关的核抗原（LANA）。 LANA在所有KS肿瘤细胞中都高度表达，并且在KSHV的生物学中起着重要作用。 LANA是潜伏期间潜在DNA复制和基因组维持所需的唯一病毒蛋白。此外，LANA与包括肿瘤抑制剂p53的多种细胞蛋白相互作用，从而调节了潜在感染的细胞中的基因表达。为了分析LANA调节细胞基因表达的能力，我们进行了基因表达分析，发现LANA调节了RB/E2F途径中的许多基因，因此，该基因可以保护淋巴样细胞免受P161NK4A诱导的细胞周期的影响。为了研究LANA在DNA复制中的作用，我们对其DNA结合特异性和活性进行了详细的生化分析。我们确定了KSHV基因组末端重复中的两个结合位点，并证明这两个位点有助于LANA支持三含质粒的复制的能力。我们假设TR内的LANA和/或顺式调节序列与促进DNA复制的细胞蛋白相互作用。为了解决这一假设，我们提出了以下三个目的，这些目的集中在解解LANA支持DNA复制的解释机制上：1）在TR中绘制复制的推定起源的顺式调节序列2）映射DNA复制的DNA和LANA依赖性DNA复制的反式依赖于lana依赖于lana的lana蛋白的lana蛋白和lana蛋白相互作用或与lana相互作用的lana和/lana rana contimal a lana-c and/appers repliqual的序列。