Regulation of cellular functions by two human Polyomaviruses

两种人类多瘤病毒对细胞功能的调节

• 批准号: 9088664
• 负责人: JAMES M PIPAS
• 金额: $ 23.1万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9088664
• 关键词: ATPase Domain; Antiviral Therapy; Binding; Binding Proteins; Biological; Biology; Capsid Proteins; Cell Nucleus; Cell Proliferation; Cell Survival; Cell physiology; Cells; Cellular biology; Cessation of life; Code; DNA Polymerase I; DNA biosynthesis; Data; Development; Disease; Family; Genetic; Genome; Goals; Growth; Hamsters; Human; Immunocompromised Host; Individual; Infection; Knowledge; Laboratories; Large T Antigen; Length; Merkel Cells; Messenger RNA; Molecular; New Jersey; Outcome; Pathology; Pathway interactions; Play; Polyomavirus; Polyomavirus Infections; Primates; Prognostic Marker; Proteins; Proteomics; RNA Splicing; Regulation; Reporting; Role; Series; Side; Signaling Protein; Simian virus 40; Site; Small T Antigen; Structure; Surface; TP53 gene; Topoisomerase; Transcriptional Regulation; Viral Tumor Antigens; Virion; Virus; base; cellular targeting; design; ds-DNA; immortalized cell; infected vector rodent; mouse polyomavirus; novel; public health relevance; senescence; transcriptome sequencing; viral DNA

 DESCRIPTION (provided by applicant): Polyomaviruses have small genomes and their coding capacity is very limited. In order to encode for functions needed in viral DNA replication, transcriptional control, and the manipulation of host functions, polyomaviruses express a series of alternatively spliced forms of their early mRNA that encode T antigens. All polyomaviruses encode a multifunctional large T antigen (LT) and most encode a small T antigen (ST), plus a series of additional poorly characterized early proteins. Of the thirteen polyomaviruses that have been isolated from humans, several express both an LT and ST that appear to be similar in function to those of the well-studied Simian virus 40 (SV40). However, other human polyomaviruses are quite distinct from SV40 regarding both the structure and biological activities of their LT and ST proteins. These include the recently discovered New Jersey polyomavirus (NJPyV) and Human polyomavirus 9 (HuPyV9). In this exploratory application we seek to uncover the unique biology of the NJPyV and HuPyV9 T antigens. Specifically, we will determine whether an alternative T antigen (AT) encoded by NJPyV is in fact an authentic middle T antigen (MT). Thus, far the only polyomaviruses that express a MT protein, murine polyomavirus and hamster polyomavirus, infect rodents. Thus, finding a human virus that expresses a MT would be very novel and suggest unique host manipulation functions. In addition, we will determine how the LT and ST from LPV, a primate polyomavirus closely related to HuPyV9- are able to independently immortalize cells and induce cell proliferation, and will also determine the molecular basis for the robust synergistic action of these proteins. Rather than recapitulate the well-trodden path that has been used to characterize T antigens in the past, we have developed new strategies designed to efficiently identify cellular pathways altered by T antigens from RNA-Seq data, and then to couple this approach with more traditional genetics and proteomics. Using this strategy, we can determine whether HuPyV9 alters the same cellular pathways as SV40 and other similar polyomaviruses, or if this virus manipulates cellular functions by novel mechanisms.

 性状（由申请方提供）：多瘤病毒基因组较小，编码能力非常有限。为了编码病毒DNA复制、转录控制和操纵宿主功能所需的功能，多瘤病毒表达一系列编码T抗原的其早期mRNA的可变剪接形式。所有多瘤病毒编码一个多功能的大T抗原（LT），大多数编码一个小T抗原（ST），加上一系列额外的特征不明显的早期蛋白质。在已经从人类分离的13种多瘤病毒中，有几种表达LT和ST，它们的功能似乎与研究充分的猿猴病毒40（SV 40）相似。然而，其他人多瘤病毒在其LT和ST蛋白的结构和生物活性方面与SV 40完全不同。其中包括最近发现的新泽西多瘤病毒（NJPyV）和人类多瘤病毒9（HuPyV 9）。在这种探索性应用中，我们试图揭示NJPyV和HuPyV 9 T抗原的独特生物学。具体而言，我们将确定由NJPyV编码的替代T抗原（AT）是否实际上是真正的中间T抗原（MT）。因此，迄今为止仅有的表达MT蛋白的多瘤病毒（鼠多瘤病毒和仓鼠多瘤病毒）感染啮齿动物。因此，发现表达MT的人类病毒将是非常新颖的，并表明独特的宿主操纵功能。此外，我们将确定LPV（一种与HuPyV 9密切相关的灵长类多瘤病毒）的LT和ST如何能够独立地使细胞永生化并诱导细胞增殖，并且还将确定这些蛋白质的强大协同作用的分子基础。我们没有重述过去用于表征T抗原的成熟途径，而是开发了新的策略，旨在从RNA-Seq数据中有效地识别T抗原改变的细胞途径，然后将这种方法与更传统的遗传学和蛋白质组学结合起来。使用这种策略，我们可以确定HuPyV 9是否改变了与SV 40和其他类似多瘤病毒相同的细胞途径，或者这种病毒是否通过新的机制操纵细胞功能。