Virology Section, Department of Life Sciences, Shiv Nadar Institution of Eminence, Gautam Buddha Nagar, Greater Noida, UP201314, India.
Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 3rd Milestone, Faridabad, Haryana, India.
Arch Virol. 2024 Oct 30;169(11):233. doi: 10.1007/s00705-024-06160-x.
Viruses have undergone evolutionary adaptations to tune their utilization of carbon sources, enabling them to extract specific cellular substrates necessary for their replication. The lack of a reliable cell culture system and a small-animal model has hampered our understanding of the molecular mechanism of replication of hepatitis E virus (HEV) genotype 1. Our recent identification of a replicative ensemble of mutant HEV RNA libraries has allowed us to study the metabolic prerequisites for HEV replication. Initial assessments revealed increased glucose and glutamine utilization during HEV replication. Inhibition of glycolysis and glycolysis + glutaminolysis reduced the levels of HEV replication to similar levels. An integrated analysis of protein-metabolite pathways suggests that HEV replication markedly alters glycolysis, the TCA cycle, and glutamine-associated metabolic pathways. Cells supporting HEV replication showed a requirement for fructose-6-phosphate and glutamine utilization through the hexosamine biosynthetic pathway (HBP), stimulating HSP70 expression to facilitate virus replication. Observations of mannose utilization and glutamine dependence suggest a crucial role of the HBP in supporting HEV replication. Inhibition of glycolysis and HSP70 activity or knockdown of glutamine fructose-6-phosphate amidotransferase expression led to a substantial reduction in HEV RNA and ORF2 expression accompanied by a significant decrease in HSP70 levels. This study demonstrates that glucose and glutamine play critical roles in facilitating HEV replication.
病毒已经进化出适应性，以调整其对碳源的利用，从而能够提取复制所需的特定细胞底物。缺乏可靠的细胞培养系统和小动物模型，阻碍了我们对戊型肝炎病毒（HEV）基因型 1 复制分子机制的理解。我们最近鉴定出复制型突变 HEV RNA 文库，使我们能够研究 HEV 复制的代谢前提条件。初步评估显示，在 HEV 复制过程中葡萄糖和谷氨酰胺的利用率增加。糖酵解和糖酵解+谷氨酰胺分解的抑制将 HEV 复制水平降低到相似水平。对蛋白质-代谢物途径的综合分析表明，HEV 复制显著改变糖酵解、三羧酸循环和与谷氨酰胺相关的代谢途径。支持 HEV 复制的细胞需要通过己糖胺生物合成途径（HBP）利用果糖-6-磷酸和谷氨酰胺，刺激 HSP70 表达以促进病毒复制。对甘露糖利用和谷氨酰胺依赖性的观察表明，HBP 在支持 HEV 复制方面起着至关重要的作用。糖酵解和 HSP70 活性的抑制或谷氨酰胺果糖-6-磷酸酰胺转移酶表达的敲低导致 HEV RNA 和 ORF2 表达显著减少，同时 HSP70 水平显著降低。这项研究表明，葡萄糖和谷氨酰胺在促进 HEV 复制方面起着关键作用。
