KSHV-encoded Small Peptites

KSHV 编码的小肽

基本信息

批准号：
8603842
负责人：
YAN YUAN
金额：
$ 20万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
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

项目摘要

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模板的相对链转录的聚腺苷酸化RNA（t3.0），并已注释为非编码RNA。 ORF50编码控制潜在和裂解生命周期之间病毒开关的复制和转录激活剂（RTA）。我们发现T3.0编码了几种小肽，其中一种（指定的病毒小肽-1或VSP-1）与RTA相互作用，并防止它通过泛素蛋白 - 蛋白酶体途径降解。结果，VSP-1促进了KSHV基因表达和裂解复制。这些发现的意义是多重的。首先，在哺乳动物和病毒基因组中，很大一部分序列被转录并注释为非编码RNA。我们的研究表明，其中一些非编码RNA可以通过编码小肽来起作用。哺乳动物细胞中可能有大量的小肽在生物过程的调节中起着关键作用。其次，我们的发现揭示了基因表达和病毒生命周期调节的新型机制。我们认为，小肽代表了重要生物学的尚未开发的来源。为了证明这一假设并估算生物学中小肽世界的大小，我们建议探索小肽对哺乳动物细胞和病毒生物学的重要性，并具有以下特定目的。（1）我们将使用蛋白质组学方法来鉴定KSHV基因组中的小肽。富集的小肽池将从KSHV感染的细胞和纯化的病毒体中制备，并进行质谱法（MS）分析。将开发特殊的潜在KSHV小肽数据库，并用于分析MS RAW光谱以鉴定KSHV小肽。（2）我们将选择一些在AIM ONE中鉴定出的代表性病毒小肽，以及两种T3.0编码的小肽（VSP-1和VSP-2），以使用遗传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，我们希望解决一个问题，是否由先前注释的非编码RNA编码的小肽是哺乳动物细胞和病毒中的常见特征，并证明了这些小肽在生物学中的生物学意义。这项研究可能导致小肽对生物调节的新领域。