UNASSIGNED: Bioinformatic analysis was conducted to identify potential homologues of RR in CyHV-2. The transcription and translation levels of ORF23 and ORF141, which showed high homology to RR, were measured during CyHV-2 replication in GICF. Co-localization experiments and immunoprecipitation were performed to investigate the interaction between ORF23 and ORF141. siRNA interference experiments were conducted to evaluate the effect of silencing ORF23 and ORF141 on CyHV-2 replication. The inhibitory effect of hydroxyurea, a nucleotide reductase inhibitor, on CyHV-2 replication in GICF cells and RR enzymatic activity in vitro was also evaluated.
UNASSIGNED: ORF23 and ORF141 were identified as potential viral ribonucleotide reductase homologues in CyHV-2, and their transcription and translation levels increased with CyHV-2 replication. Co-localization experiments and immunoprecipitation suggested an interaction between the two proteins. Simultaneous silencing of ORF23 and ORF141 effectively inhibited the replication of CyHV-2. Additionally, hydroxyurea inhibited the replication of CyHV-2 in GICF cells and the in vitro enzymatic activity of RR.
UNASSIGNED: These results suggest that the CyHV-2 proteins ORF23 and ORF141 function as viral ribonucleotide reductase and their function makes an effect to CyHV-2 replication. Targeting ribonucleotide reductase could be a crucial strategy for developing new antiviral drugs against CyHV-2 and other herpesviruses.
■进行生物信息学分析以鉴定CyHV-2中RR的潜在同源物。ORF23和ORF141的转录和翻译水平与RR有很高的同源性,在GICF中的CyHV-2复制过程中测量。进行共定位实验和免疫沉淀以研究ORF23和ORF141之间的相互作用。进行siRNA干扰实验以评估沉默ORF23和ORF141对CyHV-2复制的影响。羟基脲的抑制作用,核苷酸还原酶抑制剂,还评估了CyHV-2在GICF细胞中的复制和RR的体外酶活性。
■ORF23和ORF141被鉴定为CyHV-2中潜在的病毒核糖核苷酸还原酶同源物,它们的转录和翻译水平随CyHV-2复制而增加。共定位实验和免疫沉淀表明两种蛋白质之间存在相互作用。ORF23和ORF141的同时沉默有效地抑制了CyHV-2的复制。此外,羟基脲抑制CyHV-2在GICF细胞中的复制和RR的体外酶活性。
■这些结果表明,CyHV-2蛋白ORF23和ORF141作为病毒核糖核苷酸还原酶起作用,它们的功能对CyHV-2复制有影响。靶向核糖核苷酸还原酶可能是开发针对CyHV-2和其他疱疹病毒的新抗病毒药物的关键策略。