Lana and Cellular Gene Expression in Kaposis Sarcoma

卡波西肉瘤中的 Lana 和细胞基因表达

基本信息

批准号：
7426488
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
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-07-01 至 2011-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7426488
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

项目摘要

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）：卡波西肉瘤相关疱疹病毒 (KSHV) 也称为人类疱疹病毒 8 型 (HHV-8)，与卡波西肉瘤 (KS) 和两种淋巴组织增生性疾病相关：原发性渗出性淋巴瘤 (PEL) 及其子集多中心 Castlemen 病 (MCD)。这些恶性肿瘤的共同点是，大多数肿瘤细胞处于潜伏感染状态，仅表达少量病毒基因。其中一个基因 ORF73 编码潜伏期相关核抗原 (LANA)。 LANA 在所有 KS 肿瘤细胞中高表达，在 KSHV 生物学中发挥重要作用。 LANA 是潜伏期 DNA 复制和基因组维护所需的唯一病毒蛋白。此外，LANA 与多种细胞蛋白相互作用，包括肿瘤抑制因子 p53 和 Rb，从而调节潜伏感染细胞中的基因表达。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谱分析，发现 LANA 调节 Rb/E2F 途径中的许多基因，从而可以保护淋巴细胞免受 p161NK4a 诱导的细胞周期停滞。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。我们在 KSHV 基因组的末端重复中鉴定了两个结合位点，并证明这两个位点有助于 LANA 支持含有 TR 的质粒复制的能力。我们假设 TR 内的 LANA 和/或顺式调控序列与促进 DNA 复制的细胞蛋白相互作用。为了解决这一假设，我们提出以下三个目标，重点是破译 LANA 支持 DNA 复制的机制：1) 绘制 TR 内推定复制起点的顺式调控序列 2) 绘制 DNA 结合和 LANA 的反式要求依赖性 DNA 复制 3) 鉴定与 LANA-C 和/或最小复制起点相互作用的细胞蛋白。