Roles and regulation of polyamines during HCMV infection

多胺在 HCMV 感染过程中的作用和调节

基本信息

批准号：
10308688
负责人：
ADAM P. GEBALLE
金额：
$ 4.91万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10308688
关键词：
Affect Amino Acid Sequence Amino Acids Binding Cell physiology Cells Cistrons Codon Nucleotides Cytomegalovirus Cytomegalovirus Infections DNA Data Disease Dissection Engineering Ensure Eukaryotic Cell Eukaryotic Initiation Factors Event Experimental Designs Gene Expression Genes Genetic Human Investigation Life Link Medical Messenger RNA Metabolism Mutation Neurologic Deficit Open Reading Frames Patients Peptides Pharmacology Phenotype Physiological Polyamines Post-Translational Protein Processing Production Proline Proteins Putrescine RNA Regulation Reproduction Resistance Ribosomes Role Site Spermidine Spermine System Testing Translational Regulation Translations Viral Viral Genes Virus Virus Diseases Virus Replication alkalinity experimental study fascinate genome sequencing human pathogen immune system function in utero infant infection inhibitor insight mutant polyproline tool whole genome

项目摘要

Human cytomegalovirus (HCMV) causes life-threatening diseases in patients with poorly functioning immune systems as well as long-lasting neurological deficits in infants infected in utero. Among the many host cell factors that are needed by HCMV to replicate efficiently are small alkaline molecules called polyamines (PAs). Among their myriad activities, PAs bind to the protein synthesizing machinery in the cell. As well, they are necessary to make a protein, known as eIF5A, that is required for linking together amino acids to make certain proteins. Although it has been known for decades that the PAs are critical for HCMV to reproduce efficiently and that HCMV infection increases the intracellular concentration of PAs, the reasons why PAs are needed and the mechanisms the virus uses to ensure that there are sufficient amounts of PAs in the cells are unknown. Intriguing data emerging from studies of several host cell proteins that control the levels of PAs suggest that PAs and eIF5A might control expression of some HCMV genes, in particular a fascination one called, gpUL4. Prior studies showed that the production of gpUL4 is greatly repressed because the machinery for making it becomes trapped on the messenger RNA as a particular sequence of amino acids are being linked together. These and other results suggest that gpUL4, the function of which has been enigmatic for decades, might contribute to the control of PAs accumulation during HCMV infection. Using various HCMV mutants that affect the production of gpUL4 along with genetic and pharmacological tools for altering PA and eIF5A abundance, Aim 1 will reveal if and how PA and eIF5A alter the trapping mechanism during viral infection. Aim 2 will first test the specific hypothesis that gpUL4, alone and during HCMV infection, regulates the concentration of PAs in the cells. Another experiment will test whether HCMV can somehow adapt to reproduce well even when PA levels are low. While these studies focus on HCMV, and especially gpUL4, the results will have broad implications for understanding control of other viral and cells genes that are likely regulated by a similar mechanism.

人类巨细胞病毒 (HCMV) 会导致免疫系统功能低下的患者罹患危及生命的疾病，并导致子宫内感染的婴儿出现长期神经功能缺损。 HCMV 有效复制所需的众多宿主细胞因子中有一种称为多胺 (PA) 的碱性小分子。在其众多活性中，PA 与细胞中的蛋白质合成机制结合。此外，它们也是制造一种称为 eIF5A 的蛋白质所必需的，这种蛋白质是连接氨基酸以制造某些蛋白质所必需的。尽管数十年来人们都知道 PA 对 HCMV 有效繁殖至关重要，并且 HCMV 感染会增加细胞内 PA 浓度，但需要 PA 的原因以及病毒用于确保体内有足够量 PA 的机制细胞未知。对控制 PA 水平的几种宿主细胞蛋白的研究得出的有趣数据表明，PA 和 eIF5A 可能控制一些 HCMV 基因的表达，特别是一种名为 gpUL4 的基因。先前的研究表明，gpUL4 的产生受到极大抑制，因为当特定的氨基酸序列连接在一起时，制造 gpUL4 的机制被困在信使 RNA 上。这些和其他结果表明，几十年来其功能一直是个谜的 gpUL4 可能有助于控制 HCMV 感染期间 PA 的积累。利用影响 gpUL4 产生的各种 HCMV 突变体以及改变 PA 和 eIF5A 丰度的遗传和药理学工具，目标 1 将揭示 PA 和 eIF5A 是否以及如何改变病毒感染期间的捕获机制。目标 2 将首先测试 gpUL4 单独和在 HCMV 感染期间调节细胞中 PA 浓度的具体假设。另一项实验将测试 HCMV 是否能够在 PA 水平较低的情况下以某种方式适应并良好繁殖。虽然这些研究集中于 HCMV，尤其是 gpUL4，但结果将对理解可能受类似机制调节的其他病毒和细胞基因的控制产生广泛的影响。