APOBEC3G and RNA granules: restricting HIV infection and Alu retrotransposition

APOBEC3G 和 RNA 颗粒：限制 HIV 感染和 Alu 逆转座

基本信息

批准号：
7339117
负责人：
Ya-Lin Chiu
金额：
$ 16.2万
依托单位：
J. DAVID GLADSTONE INSTITUTES
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-08 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7339117
关键词：
AIDS/HIV problem Acute Myelocytic Leukemia Affect Alu Elements Anti-Retroviral Agents Antiviral Agents Binding Biochemical Biological Biological Assay Biology CD4 Positive T Lymphocytes Cancer cell line Cell Line Cells Characteristics Class Complex Cytoplasmic Granules DNA Dendritic Cells Development Disease Electrophoretic Mobility Shift Assay Elements Enzymes Event Family Frequencies Genes Genetic Genetic Transcription Genome Genomic Instability Genotoxic Stress Goals Growth HIV HIV Infections HIV-1 Hereditary Disease Human Human Activities Human Genome Image Immunity Infection Control Lead Leukemic Cell Life Link Location Long Interspersed Nucleotide Elements Malignant Neoplasms Mediating Messenger RNA Methods Mobile Genetic Elements Molecular Molecular Profiling Mutagenesis Mutation Nuclear Nucleic Acid Regulatory Sequences Nucleosides Pattern Physiological Play Polymerase Chain Reaction Polyribosomes Principal Investigator Process Property Proteins Publications RNA RNA Binding RNA-Directed DNA Polymerase Radiation therapy Range Recruitment Activity Research Research Personnel Rest Retroelements Retrotransposition Retroviridae Reverse Transcription Ribonucleoproteins Role SS-A antigen Screening procedure Second Primary Cancers Series Signal Transduction Site Small Interfering RNA Solid Stress Structure Surveys T-Lymphocyte Testing Therapeutic Time Translations Treatment-Related Cancer Untranslated Regions Viral Proteins Viral Reverse Transcription Virion acrosome stabilizing factor alpha-Thalassemia apolipoprotein B mRNA editing enzyme base cancer cell cancer therapy cellular imaging chemotherapy cofactor deoxycytidine deaminase endonuclease human CEM15 protein in vivo insight knock-down leukemia loss of function molecular mass monocyte multicatalytic endopeptidase complex mutant neoplastic novel strategies peripheral blood polypeptide prevent programs response trafficking

项目摘要

DESCRIPTION (provided by applicant): The discovery of human APOBEC3G (A3G) and related deoxycytidine deaminases as potent intrinsic anti-HIV factors represented an exciting advance in the field of HIV/AIDS research. The later discovery that HIV Vif counters the antiviral effects of A3G by targeting this factor for rapid proteasome-mediated degradation has spawned efforts to create a new class of HIV therapeutics. New insights into the rich biology of A3G have also been gained, including the observation that A3G is expressed in different forms in different cells. Resting CD4 T cells express a low-molecular-mass (LMM) form of A3G that actively restricts the growth of HIV. Conversely, activated CD4 T cells express a high-molecular-mass (HMM) form of A3G that no longer restricts HIV but instead interrupts the retrotransposition of endogenous retroelements like Alu. Intriguingly, Staufen-containing RNA granules form a major component of the HMM A3G complexes. I hypothesize that the biology of A3G is inextricably linked to the inducible formation, dynamic spatial localization, and controlled disassembly of these RNA granules. To test this hypothesis, I propose three specific aims. In Aim 1, I will study the intracellular signals and molecular interactions that govern the formation and disassembly of A3G containing RNA granules and decipher their potential roles in regulating the expression and antiviral activity of human A3G. In Aim 2, I will explore how A3G induces the recruitment of Alu RNA into these RNA granules and in turn how A3G regulates Alu retrotransposition. I will also assess how these interactions modulate the intrinsic antiviral activity of A3G. In Aim 3, I will test the potential role of A3G in controlling Alu-mediated genomic instability and the contribution of such instability to the development of secondary leukemias occurring after genotoxic cancer therapy. For these studies, I will utilize biochemical methods, genome-wide siRNA screening, molecular beacon- and FRET-based imaging approaches in both primary T cells and leukemic cells. The proposed studies promise to provide new insights into the previously unrecognized role of A3G complexes containing RNA granules in controlling retrotransposition of endogenous retroelements. The findings will expand our understanding of A3G biology and could ultimately lead to novel strategies both to protect activated CD4 T cells from HIV infection and to minimize the frequency of cancer therapy-related secondary leukemias.

描述（由申请人提供）：人类 APOBEC3G (A3G) 和相关脱氧胞苷脱氨酶作为有效的内在抗 HIV 因子的发现代表了 HIV/AIDS 研究领域的令人兴奋的进展。后来发现 HIV Vif 通过针对 A3G 因子进行蛋白酶体介导的快速降解来对抗 A3G 的抗病毒作用，这引发了创建新型 HIV 疗法的努力。还获得了对 A3G 丰富生物学的新见解，包括观察到 A3G 在不同细胞中以不同形式表达。静息 CD4 T 细胞表达低分子质量 (LMM) 形式的 A3G，可主动限制 HIV 的生长。相反，激活的 CD4 T 细胞表达高分子量 (HMM) 形式的 A3G，它不再限制 HIV，而是中断 Alu 等内源性逆转录因子的逆转录转座。有趣的是，含有诗道芬的 RNA 颗粒形成了 HMM A3G 复合物的主要成分。我假设 A3G 的生物学特性与这些 RNA 颗粒的诱导形成、动态空间定位和受控分解有着千丝万缕的联系。为了检验这个假设，我提出了三个具体目标。在目标 1 中，我将研究控制含有 RNA 颗粒的 A3G 形成和分解的细胞内信号和分子相互作用，并破译它们在调节人类 A3G 表达和抗病毒活性中的潜在作用。在目标 2 中，我将探讨 A3G 如何诱导 Alu RNA 募集到这些 RNA 颗粒中，以及 A3G 如何调节 Alu 逆转录转座。我还将评估这些相互作用如何调节 A3G 的内在抗病毒活性。在目标 3 中，我将测试 A3G 在控制 Alu 介导的基因组不稳定性方面的潜在作用，以及这种不稳定性对基因毒性癌症治疗后发生的继发性白血病发展的贡献。对于这些研究，我将在原代 T 细胞和白血病细胞中利用生化方法、全基因组 siRNA 筛选、分子信标和基于 FRET 的成像方法。拟议的研究有望为含有 RNA 颗粒的 A3G 复合物在控制内源性反转录元件的反转录转座中的先前未被认识的作用提供新的见解。这些发现将扩大我们对 A3G 生物学的理解，并最终可能产生新的策略，既保护活化的 CD4 T 细胞免受 HIV 感染，又最大限度地减少癌症治疗相关的继发性白血病的发生率。