Human cellular factors restricting HIV replication

限制HIV复制的人体细胞因素

基本信息

批准号：
8055454
负责人：
Susana T Valente
金额：
$ 10.8万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-05 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8055454
关键词：
26S proteasome Animals Antiviral Agents Applied Genetics Binding Bioterrorism Cell Line Cell Nucleus Cells Chimeric Proteins Clinical Complementary DNA Complex Cytoplasm Epithelial Cells Eukaryotic Initiation Factor-3 Family Flaviviridae Gene Library Genes Genetic Screening Genome Guanosine Triphosphate HIV HIV Infections HIV-1 HIV-2 Hepatitis C virus Homologous Gene Human Human Virus Individual Infection Infection prevention Integration Host Factors Interferons Investigation Knowledge Lead Libraries Life Cycle Stages Long Terminal Repeats Mammalian Cell Maps Mediating Medical Messenger RNA Morbidity - disease rate Mouse Protein N-terminal Nature Nuclear Nuclear Export Peptide Library Peptides Peripheral Blood Lymphocyte Play Population Primates Protein Binding Proteins RNA RNA Binding RNA Processing RNA Viruses RNA, Transfer, Met Resistance Resistance to infection Retroviral Vector Retroviridae Role SIV Screening procedure Testing Translations Viral Virus Virus Diseases Virus Replication Yellow fever virus Zeocin combat design in vivo interest mRNA Export mRNA Expression macrophage mortality novel public health relevance research study response therapeutic target vector

项目摘要

DESCRIPTION (provided by applicant): Cellular proteins are now appreciated as critically involved in all steps of the human immunodeficiency virus type 1 (HIV-1) life cycle, and disrupting host functions essential for virus replication may provide novel antiviral approaches. To identify new molecules in human cells critical for blocking HIV-1 replication we established a genetic screen of mammalian complementary DNAv(cDNA) libraries for genes that prevent infection by a genetically marked retrovirus. A cDNA library from human cells (HeLa) stimulated by interferon (known to activate many antiviral proteins) was generated, introduced into a human virus-susceptible cell line, and a virus resistant population was selected for survival after multiple rounds of infection with an HIV vector expressing a toxic gene. Two active cDNAs were identified, dubbed H1 and H2: H1 expresses the N- terminal portion (86a.a.) of heterogeneous nuclear ribonuclear protein U (hnRNP U) and H2 the N-terminal portion (93 a.a.) of the eukaryotic initiation factor 3 subunit p47 (elF3p47 or elF3f). hnRNP U is involved in pre RNA processing, mRNA transport to the cytoplasm, intracellular localization, and turnover of mRNAs. elF3f belongs to the elF3 complex that plays a role in translation by associating elF2-GTP-Met-tRNA and promoting RNA binding. elF3f is also related to a mouse protein called Mov34; the human homologue of Mov34 is the S12 subunit of the 26S proteasome. Both cDNAs were characterized as necessary and sufficient to confer resistance to HIV infection. However, H1 expressing cells seems to induce retention of the viral mRNA in the nucleus, while H2 expressing cells induce a drastic reduction in the amount of the nuclear viral mRNA by specifically reducing cleavage activity. In both cases levels of the viral messages in the cytoplasm are dramatically reduced. Both gene fragments specifically target the 3' long terminal repeat 3'LTR) in the viral mRNA. These results suggest that HIV-1 requires machinery for the nuclear export/cleavage of the mRNA message that can be specifically inhibited by the identified cDNAs. We propose to continue and expand these investigations, as well as further identify more interfering factors by screening cDNA, peptide or RNA interfering (RNAi) libraries. The discovery of cellular factors involved in retroviral replication and an increased knowledge of their mode of action may lead to important antiviral approaches in clinical settings.

描述（由申请人提供）：细胞蛋白现在被认为关键参与1型人类免疫缺陷病毒（HIV-1）生命周期的所有步骤，并且破坏病毒复制所必需的宿主功能可能提供新的抗病毒方法。为了识别人类细胞中对阻断 HIV-1 复制至关重要的新分子，我们建立了哺乳动物互补 DNAv(cDNA) 文库的遗传筛选，以寻找预防基因标记逆转录病毒感染的基因。产生干扰素（已知可激活许多抗病毒蛋白）刺激的人类细胞（HeLa）的 cDNA 文库，将其引入人类病毒敏感细胞系，并选择病毒抗性群体在 HIV 多轮感染后存活表达有毒基因的载体。鉴定出两个活性 cDNA，分别称为 H1 和 H2：H1 表达异质核核糖核蛋白 U (hnRNP U) 的 N 端部分 (86 个氨基酸)，H2 表达真核起始因子的 N 端部分 (93 个氨基酸) 3 个亚基 p47（elF3p47 或 eF3f）。 hnRNP U 参与前 RNA 加工、mRNA 转运至细胞质、细胞内定位和 mRNA 周转。 eF3f属于elF3复合体，通过缔合elF2-GTP-Met-tRNA并促进RNA结合而在翻译中发挥作用。 eF3f 还与一种名为 Mov34 的小鼠蛋白有关； Mov34 的人类同源物是 26S 蛋白酶体的 S12 亚基。两种 cDNA 都被认为是赋予对 HIV 感染抵抗力所必需和充分的。然而，H1 表达细胞似乎诱导病毒 mRNA 保留在细胞核中，而 H2 表达细胞则通过特异性降低裂解活性，诱导核病毒 mRNA 量急剧减少。在这两种情况下，细胞质中的病毒信息水平都显着降低。两个基因片段都特异性靶向病毒 mRNA 中的 3' 长末端重复序列（3'LTR）。这些结果表明，HIV-1 需要用于 mRNA 信息的核输出/切割的机制，该机制可以被已识别的 cDNA 特异性抑制。我们建议继续并扩大这些研究，并通过筛选 cDNA、肽或 RNA 干扰 (RNAi) 文库进一步鉴定更多干扰因素。参与逆转录病毒复制的细胞因子的发现以及对其作用模式的了解的增加可能会导致临床环境中重要的抗病毒方法。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Somatic cell genetic analyses to identify HIV-1 host restriction factors.

体细胞遗传分析以确定 HIV-1 宿主限制因素。

DOI：
发表时间：
2009
期刊：
影响因子：
0
作者：
Valente, Susana T;Goff, Stephen P
通讯作者：
Goff, Stephen P

Inhibition of HIV-1 gene expression by a fragment of hnRNP U.

hnRNP U 片段抑制 HIV-1 基因表达。

DOI：
10.1016/j.molcel.2006.07.021
发表时间：
2006-08-18
期刊：
Molecular cell
影响因子：
16
作者：
S. Valente;S. Goff
通讯作者：
S. Goff

Inhibition of HIV-1 replication by eIF3f.

eIF3f 抑制 HIV-1 复制。

DOI：
发表时间：
2009-03-17
期刊：
影响因子：
11.1
作者：
Valente, Susana T;Gilmartin, Greg M;Mott, Christina;Falkard, Brie;Goff, Stephen P
通讯作者：
Goff, Stephen P

An analog of the natural steroidal alkaloid cortistatin A potently suppresses Tat-dependent HIV transcription.

天然甾体生物碱 Cortistatin A 的类似物可有效抑制 Tat 依赖性 HIV 转录。

DOI：
10.1016/j.chom.2012.05.016
发表时间：
2012-07-19
期刊：
Cell host & microbe
影响因子：
30.3
作者：
Mousseau G;Clementz MA;Bakeman WN;Nagarsheth N;Cameron M;Shi J;Baran P;Fromentin R;Chomont N;Valente ST
通讯作者：
Valente ST

HIV-1 mRNA 3' end processing is distinctively regulated by eIF3f, CDK11, and splice factor 9G8.

HIV-1 mRNA 3 末端加工明显受到 eIF3f、CDK11 和剪接因子 9G8 的调节。