Studying the role of KSHV-encoded microRNAs

研究 KSHV 编码的 microRNA 的作用

基本信息

批准号：
7753243
负责人：
ROLF F RENNE
金额：
$ 29.71万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-06 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7753243
关键词：
Address Angiogenesis Inhibitors Apoptosis Binding Bioinformatics Biological Biological Assay Biological Process Biology Cell Proliferation Cells Cultured Cells DNA Viruses Data Detection Endothelial Cells Environment Epithelial Cells Escherichia coli Functional RNA Gene Expression Gene Expression Profiling Gene Targeting Genes Genomics Goals Hematopoiesis Herpesviridae Human Human Herpesvirus 4 Human Herpesvirus 8 Infection Journals Kaposi Sarcoma Kinetics Knock-out Laboratories Length Life Cycle Stages Locales Lymphoid Lymphoma Lytic Mammals Maps MicroRNAs Multicentric Angiofollicular Lymphoid Hyperplasia Northern Blotting Organism Pathogenesis Pathway interactions Plants Play Post-Transcriptional Regulation RNA Recombinants Regulator Genes Reporting Reverse Transcriptase Polymerase Chain Reaction Role Sampling Sequence Analysis Small RNA Technology Testing Thrombospondin 1 Tissue Sample Transcript Translations Untranslated Regions Vero Cells Viral Viral Genes Viral Genome Virus Work base effusion gammaherpesvirus interest lytic replication mRNA Transcript Degradation novel pathogen prevent programs promoter recombinant virus transcription factor tumor tumorigenesis virology

项目摘要

DESCRIPTION (provided by applicant): MicroRNAs are small non-coding regulatory RNA molecules that bind to 3'UTRs of mRNAs to either prevent their translation or induce their degradation. Previously identified in a variety of organisms ranging from plants to mammals, miRNAs are also now known to be produced by DNA viruses. The human ?-herpesvirus Epstein-Barr Virus has been shown to encode miRNAs which potentially regulate both viral and cellular genes. To determine whether Kaposi's Sarcoma-associated herpesvirus (KSHV) encodes miRNAs, we cloned small RNAs from KSHV positive primary effusion lymphoma derived cells. Sequence analysis revealed 11 isolated RNAs of 19 to 23 bases in length that perfectly align to KSHV. Surprisingly, all candidate miRNAs mapped to a single genomic locale within the latency-associated region of KSHV (Samols et al., Journal of Virology 2005). While our work was in review, two other laboratories reported the identification of KSHV-encoded miRNAs. Hence, these data suggest that virally-encoded miRNAs represent a novel mechanism by which KSHV may regulate viral and/or cellular gene expression during both latent and lytic KSHV replication. In human cells, over 450 miRNAs have been identified. Targets and functions of only a few miRNAs have been experimentally determined thus far, yet some miRNAs such as human hsa-miR-14 and hsa-miR-181 regulate fundamental biological processes like apoptosis, cell proliferation, and hematopoiesis and very recently have been implicated in tumorigenesis. Based on our preliminary results we hypothesize that miRNAs play an important role in the KSHV life cycle and may contribute to KSHV pathogenesis. To directly address this hypothesis we propose the following specific aims: SA1: Analysis of KSHV miRNA expression during latent and lytic replication in cultured cells and tissues samples of different origins. SA2: Identification of cellular and viral targets regulated by KSHV-encoded miRNAs. SA3: Evaluate the role of miRNA expression in the context of the viral genome. In support of our hypothesis we provide new preliminary data on identifying cellular miRNA targets in stably miRNA-expressing 293 cells (Samols et al., PLoS Pathogens, 2007, May 11). In summary, the identification of virally encoded miRNAs within the latency-associated region of KSHV is novel and very exciting as it suggests an entirely new level of regulating gene expression by which KSHV can potentially reprogram the host cell environment.

描述（由申请人提供）：microRNA是小的非编码调节RNA分子，与3'UTRS结合mRNA，以防止其翻译或诱导其降解。先前在从植物到哺乳动物的各种生物中鉴定出的miRNA现在也已知由DNA病毒产生。人类的Hepesvirus Epstein-Barr病毒已被证明可以编码可能调节病毒和细胞基因的miRNA。为了确定Kaposi的肉瘤相关疱疹病毒（KSHV）是否编码miRNA，我们从KSHV阳性原发性积液淋巴瘤衍生的细胞中克隆了小RNA。序列分析表明，11至23个碱基的11个孤立RNA与KSHV完全一致。令人惊讶的是，所有候选miRNA都映射到KSHV潜伏相关区域内的单个基因组部位（Samols等人，病毒学杂志，2005年）。当我们的工作正在审查时，另外两个实验室报告了KSHV编码的miRNA的识别。因此，这些数据表明，病毒编码的miRNA代表了一种新的机制，在潜在和裂解KSHV复制过程中，KSHV可以调节病毒和/或细胞基因表达。在人类细胞中，已经鉴定出超过450个miRNA。到目前为止，仅在实验中确定了仅几个miRNA的靶和功能，但是一些miRNA（例如人类HSA-MIR-14和HSA-MIR-181）调节基本生物学过程，例如凋亡，细胞增殖和造血以及最近与肿瘤发生有关。根据我们的初步结果，我们假设miRNA在KSHV生命周期中起着重要作用，并且可能有助于KSHV发病机理。为了直接解决这一假设，我们提出了以下特定目的： SA1：分析不同起源的培养细胞和组织样品中潜在和裂解复制过程中的KSHV miRNA表达。 SA2：鉴定由KSHV编码的miRNA调节的细胞和病毒靶标。 SA3：评估miRNA表达在病毒基因组背景下的作用。为了支持我们的假设，我们提供了有关识别稳定表达miRNA的293个细胞中细胞miRNA靶标的新初步数据（Samols等，Plos病原体，2007年5月11日）。总而言之，在KSHV的潜伏相关区域内鉴定了病毒编码的miRNA是新颖的，非常令人兴奋，因为它表明了一个全新的调节基因表达水平，KSHV可以通过该水平可以通过该水平来重新编程宿主细胞环境。