The identification of viral ribonucleotide reductase encoded by ORF23 and ORF141 genes and effect on CyHV-2 replication

Wenjie Cheng; Wenjie Cheng; Wenjie Cheng; Qikang Chen; Qikang Chen; Qikang Chen; Yilin Ren; Yilin Ren; Yilin Ren; Ye Zhang; Ye Zhang; Ye Zhang; Ye Zhang; Liqun Lu; Liqun Lu; Liqun Lu; Liqun Lu; Lang Gui; Lang Gui; Lang Gui; Lang Gui; Dan Xu; Dan Xu; Dan Xu; Dan Xu

doi:10.3389/fmicb.2023.1154840

Frontiers in Microbiology (Apr 2023)

The identification of viral ribonucleotide reductase encoded by ORF23 and ORF141 genes and effect on CyHV-2 replication

Wenjie Cheng,
Wenjie Cheng,
Wenjie Cheng,
Qikang Chen,
Qikang Chen,
Qikang Chen,
Yilin Ren,
Yilin Ren,
Yilin Ren,
Ye Zhang,
Ye Zhang,
Ye Zhang,
Ye Zhang,
Liqun Lu,
Liqun Lu,
Liqun Lu,
Liqun Lu,
Lang Gui,
Lang Gui,
Lang Gui,
Lang Gui,
Dan Xu,
Dan Xu,
Dan Xu,
Dan Xu

Affiliations

Wenjie Cheng: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Wenjie Cheng: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Wenjie Cheng: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Qikang Chen: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Qikang Chen: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Qikang Chen: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Yilin Ren: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Yilin Ren: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Yilin Ren: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Ye Zhang: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Ye Zhang: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Ye Zhang: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Ye Zhang: National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China
Liqun Lu: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Liqun Lu: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Liqun Lu: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Liqun Lu: National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China
Lang Gui: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Lang Gui: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Lang Gui: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Lang Gui: National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China
Dan Xu: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Dan Xu: National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China
Dan Xu: Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China
Dan Xu: National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China

DOI: https://doi.org/10.3389/fmicb.2023.1154840
Journal volume & issue: Vol. 14

Abstract

Read online

IntroductionRibonucleotide reductase (RR) is essential for the replication of the double-stranded DNA virus CyHV-2 due to its ability to catalyze the conversion of ribonucleotides to deoxyribonucleotides, and is a potential target for the development of antiviral drugs to control CyHV-2 infection.MethodsBioinformatic 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.ResultsORF23 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.ConclusionThese 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.

Published in Frontiers in Microbiology

ISSN: 1664-302X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: http://www.frontiersin.org/journals/microbiology

About the journal

Abstract

Keywords