Viral RNPs, mRNA Stability and Export

病毒 RNP、mRNA 稳定性和输出

• 批准号: 8307755
• 负责人: JOAN A. STEITZ
• 金额: $ 18.78万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8307755
• 关键词: 3' Untranslated Regions; Antibodies; Antinuclear Antibodies; B-Lymphocytes; Binding; Binding Proteins; Biological Assay; Callithrix; Cancer Etiology; Cebidae; Cell Line; Cell Nucleus; Cells; Collaborations; Complex; Elements; Epstein-Barr virus encoded RNA 1; Epstein-Barr virus encoded RNA 2; Exhibits; Functional RNA; Gene Expression; Growth; Herpesviridae; Homologous Gene; Human; Human Herpesvirus 4; Human Herpesvirus 8; Immunocompromised Host; Immunofluorescence Immunologic; In Vitro; Individual; Infection; Infectious Mononucleosis; Left; Length; Luciferases; Lymphoid Cell; Lymphoma; Lytic; Lytic Phase; Malignant Neoplasms; Messenger RNA; Metabolism; MicroRNAs; Molecular; Monkeys; NMR Spectroscopy; Nuclear; Nuclear RNA; Nucleotides; Pathway interactions; Phenotype; Poly(A)+ RNA; Primates; Proteins; RNA; RNA Decay; RNA Splicing; Reporter; Repression; Resistance; Resolution; Ribonucleoproteins; Roentgen Rays; Role; Saimiriine Herpesvirus 2; Site; Small RNA; Structure; T-Lymphocyte; Tail; Testing; Therapeutic; Translational Repression; Translations; Untranslated Regions; Viral; Viral Physiology; Virus; X-Ray Crystallography; antigen binding; base; cell transformation; combat; design; gammaherpesvirus; gene function; in vivo; insight; leukemia/lymphoma; mRNA Export; mRNA Stability; metaplastic cell transformation; novel; prevent; sarcoma; translation factor; viral RNA

The roles of non-coding RNAs in lymphoid cells harboring three transforming herpesviruses are being investigated. Epstein-Barr virus (EBV) infects and transforms humanB cells; it is the causative agent of infectious mononucleosis and is associated with several human cancers. Herpesvirus saimiri (HVS) induces fatal lymphomas and leukemias in New World monkeys and transforms human and monkey T lymphocytes in culture, generating a mature; activated phenotype. Kaposi's sarcoma-associated herpesvirus (KSHV) afflicts immunocompromised individuals and persists in a latent form until lytic activation. The two EBVencoded EBERs, as well as >23 newly identified microRNAs, and the seven HVS-encoded HSURs are expressed in virally transformed cells. Upon induction, KSHV produces PAN, a capped, polyadenylated RNA that does not leave the nucleus. These viral RNAs are all small (from 21 nts to 1.1 kb), abundant, conserved, and associate with host proteins to form RNPs. Studying their functions has,already contributed important insights into host cell pathways perturbed during viral transformation or lytic growth. Proposed aims will test the hypotheses that EBERs, HSURs and PAN manipulate cellular metabolism by sequestering host proteins or microRNAs, or that the resulting RNPs perform a critical viral function. We recently found that HSUR RNPs bind certain host microRNAs; interference with their regulatory roles in transformed T cells will be examined. The possibility that EBERs likewise bind host microRNAs will be tested. Our discovery that microRNAs in quiescent cells activate rather than repress translation will be exploited to investigate the EBV-encoded microRNAs since many transformed B cells exist in a quiescent state in the body. We showed that PAN RNA accumulates to very high nuclear levels in KSHV-reactivated cells because an element (the ENE) near its 3' end engages the polyA tail to prevent RNA decay. Structural studies of the ENE +/- oligoA, as well as complexed with polyA binding protein (PABPC1), will provide mechanistic insights. Our observation that PAN RNA is involved in relocalization of host PABPC1 to the nucleus, suggesting collaboration with the KSHV SOX protein in host mRNA shut off, will be investigated

非编码RNA在携带三种转化型疱疹病毒的淋巴细胞中的作用正在研究中。 研究了EB病毒（EBV）感染并转化人类B细胞;它是EB病毒感染的病原体。 传染性单核细胞增多症，并与几种人类癌症有关。松鼠猴疱疹病毒（HVS）诱导 新世界猴的致命性淋巴瘤和白血病，并转化人类和猴的T淋巴细胞 在培养中，产生成熟的;活化的表型。卡波西肉瘤相关疱疹病毒 折磨免疫功能低下的个体，并以潜伏形式持续存在，直到裂解活化。两个EB病毒编码的 EBER，以及>23个新发现的microRNA，以及7个HVS编码的HSUR， 在病毒转化的细胞中表达。在诱导下，KSHV产生PAN，一种加帽的多腺苷酸化RNA 不会离开细胞核。这些病毒RNA都很小（从21 nt到1.1 kb），丰富，保守， 并与宿主蛋白质结合形成RNP。研究它们的功能， 深入了解病毒转化或裂解生长期间干扰的宿主细胞途径。 提出的目标将检验EBER、HSUR和PAN通过以下方式操纵细胞代谢的假设： 隔离宿主蛋白质或微小RNA，或者产生的RNP执行关键的病毒功能。我们 最近发现，HSUR RNP结合某些宿主microRNA;干扰它们的调节作用， 将检查转化的T细胞。EBER同样结合宿主microRNA的可能性将是 测试.我们发现，在静止细胞中的microRNA激活而不是抑制翻译， 由于许多转化的B细胞存在于静止的细胞中， 身体里的状态。我们发现PAN RNA在KSHV再激活的细胞核中积累到非常高的水平， 因为在其3'端附近的元件（ENE）接合polyA尾以防止RNA衰变。结构 ENE +/- oligoA以及与polyA结合蛋白（PABPC 1）复合的研究将提供 机械的洞察力。我们观察到PAN RNA参与了宿主PABPC 1的重新定位， 细胞核，表明与KSHV SOX蛋白在宿主mRNA关闭中的协作，将被研究