Viral RNPs, mRNA Stability and Export

病毒 RNP、mRNA 稳定性和输出

基本信息

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
项目类别：
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&apos 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

项目摘要

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

携带三种转化疱疹病毒的淋巴细胞中非编码 RNA 的作用正在被研究调查了。 Epstein-Barr 病毒 (EBV) 感染并转化人类 B 细胞；它是传染性单核细胞增多症，与多种人类癌症有关。赛米尔疱疹病毒 (HVS) 诱导新大陆猴致命性淋巴瘤和白血病，并转化人类和猴 T 淋巴细胞在文化上，生成成熟；激活的表型。卡波西肉瘤相关疱疹病毒 (KSHV) 影响免疫功能低下的个体，并以潜伏形式持续存在，直至裂解激活。两个 EBVencoded EBER 以及 > 23 个新识别的 microRNA 和 7 个 HVS 编码的 HSUR 在病毒转化的细胞中表达。诱导后，KSHV 产生 PAN，一种加帽的聚腺苷酸化 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、并与宿主蛋白结合形成 RNP。研究它们的功能已经做出了重要贡献深入了解病毒转化或裂解生长过程中宿主细胞途径的扰动。拟议的目标将测试 EBER、HSUR 和 PAN 通过以下方式操纵细胞代谢的假设：隔离宿主蛋白或 microRNA，或者由此产生的 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；干扰其监管作用将检查转化的 T 细胞。 EBER 同样结合宿主 microRNA 的可能性是已测试。我们发现静止细胞中的 microRNA 会激活而不是抑制翻译由于许多转化的 B 细胞存在于静止状态，因此可用于研究 EBV 编码的 microRNA 体内的状态。我们发现 PAN RNA 在 KSHV 重新激活的细胞核中积累到非常高的水平细胞，因为其 3' 端附近的一个元件（ENE）与 PolyA 尾部接合以防止 RNA 衰变。结构性 ENE +/- 寡A以及与多聚A结合蛋白（PABPC1）复合的研究将提供机制的见解。我们观察到 PAN RNA 参与宿主 PABPC1 重新定位到细胞核，表明与 KSHV SOX 蛋白在宿主 mRNA 关闭中的合作，将进行研究