KSHV-encoded Small Peptites

KSHV 编码的小肽

• 批准号: 8542364
• 负责人: YAN YUAN
• 金额: $ 23.24万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-15 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8542364
• 关键词: Address; Biological; Biological Process; Biology; Categories; Cells; Computer software; DNA; Databases; Expeditions; Functional RNA; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genetic; Genetic Transcription; Genome; Human Herpesvirus 8; Intercistronic Region; Investigation; Lead; Life Cycle Stages; Lytic; Mammalian Cell; Mediating; MicroRNAs; Molecular Virology; Open Reading Frames; Pathway interactions; Peptides; Play; Poly(A)+ RNA; Proteomics; RNA; Recombinants; Regulation; Research; Role; Source; Transcription Coactivator; Ubiquitin; Untranslated RNA; Viral; Viral Genome; Virion; Virus; genome-wide; genome-wide analysis; lytic replication; mammalian genome; multicatalytic endopeptidase complex; novel; prevent; public health relevance

DESCRIPTION (provided by applicant): A polyadenylated RNA of 3.0 kb (T3.0) is transcribed from the opposite strand of ORF50 DNA template in the KSHV genome and has been annotated as a non-coding RNA. ORF50 encodes the replication and transcription activator (RTA) that controls switch of the virus between latent and lytic life cycle. We found that T3.0 encodes several small peptides and one of them (designated viral Small Peptide-1 or vSP-1) interacts with RTA and prevents it from being degraded through the ubiquitin-proteasome pathway. As a consequence, vSP-1 facilitates KSHV gene expression and lytic replication. The significance of these findings is multi-fold. First, in both mammalian and viral genomes, a large proportion of sequences are transcribed and annotated as noncoding RNAs. Our study suggests that some of these noncoding RNAs may function through encoding small peptides. There might be large numbers of small peptides in mammalian cells that play critical roles in regulation of biological processes. Second, our finding revealed a novel mechanism of regulation of gene expression as well as viral life cycle. We believe that small peptides represent an untapped source of important biology. To prove this hypothesis and to estimate the magnitude of small peptide world in biology, we proposed to explore the significance of small peptides to biology of mammalian cells and viruses with the specific aims as follows. (1) We would use proteomics approach to identify small peptides from the KSHV genome. Enriched small peptide pools will be prepared from KSHV-infected cells as well as purified virions and subjected to mass spectrometric (MS) analysis. A special potential KSHV small peptide database will be developed and used to analyze the MS raw spectra for identification of KSHV small peptides. (2) We will choose a few representative viral small peptides identified in aim one, together with two T3.0-encoded small peptides (vSP-1 and vSP- 2), to assess the functional roles of viral small peptides in KSHV life cycle using genetic approaches. Through this exploratory investigation, we hope to address the question if small peptides encoded by previously annotated noncoding RNAs are common features in mammalian cells and viruses and to demonstrate the biological significance of these small peptides in biology. This study may lead to a new field on biological regulation by small peptides.

描述（由申请人提供）：从KSHV基因组中ORF50 DNA模板的对链转录出一个3.0 kb （T3.0）的聚腺苷化RNA，并被注释为非编码RNA。ORF50编码复制和转录激活子（RTA），控制病毒在潜伏和裂解生命周期之间的切换。我们发现T3.0编码了几个小肽，其中一个（称为病毒小肽-1或vSP-1）与RTA相互作用，并通过泛素-蛋白酶体途径阻止RTA被降解。因此，vSP-1促进了KSHV基因的表达和裂解复制。这些发现的意义是多方面的。首先，在哺乳动物和病毒基因组中，大部分序列被转录和注释为非编码rna。我们的研究表明，其中一些非编码rna可能通过编码小肽发挥作用。哺乳动物细胞中可能存在大量的小肽，它们在调节生物过程中起着关键作用。其次，我们的发现揭示了基因表达和病毒生命周期调控的新机制。我们相信小肽代表了重要的生物学尚未开发的来源。为了证明这一假设，估计小肽世界在生物学中的重要性，我们提出探索小肽对哺乳动物细胞和病毒生物学的意义，具体目的如下：(1)我们将使用蛋白质组学方法从KSHV基因组中鉴定小肽。从感染kshv的细胞和纯化的病毒粒子中制备富集的小肽池，并进行质谱分析。我们将建立一个特殊的潜在KSHV小肽数据库，用于分析KSHV小肽的质谱。(2)我们将选择目标1中鉴定的几个具有代表性的病毒小肽，以及两个t3.0编码的小肽（vSP-1和vSP- 2），利用遗传学方法评估病毒小肽在KSHV生命周期中的功能作用。通过这项探索性研究，我们希望解决由先前注释的非编码rna编码的小肽是否为哺乳动物细胞和病毒的共同特征的问题，并证明这些小肽在生物学上的生物学意义。该研究可能为小肽生物调控开辟新的研究领域。