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Published on: September 27, 2014
Establishment of minigenomes for infectious bursal disease virus
Hui Yang1,2, Mingrui Zhang3, Sanying Wang4
1Disease Intervention and Prevention Program, Texas Biomedical Research Institute, San Antonio, TX, USA. huiy@yzu.edu.cn.
Abstract:
Minigenomes (MGs) have greatly advanced research on the viral life cycle, including viral replication and transcription, virus‒host interactions, and the discovery of antivirals against RNA viruses. However, an MG for infectious bursal disease virus (IBDV) has not been well established. Here, we describe the development of IBDV MG, in which the entire coding sequences of viral genomic segments A and B are replaced with Renilla luciferase (Rluc) or enhanced green fluorescent protein (EGFP) reporter genes. Under the control of the RNA polymerase I promoter, the translation of IBDV MG is controlled by the viral proteins VP1 and VP3. Interestingly, IBDV B MG shows greater activity than does IBDV A MG. Moreover, the sense IBDV B MG was expressed at a higher level than the antisense IBDV B MG. In agreement with our previous findings, the translation of IBDV B MG controlled by VP1 and VP3 is independent of the cellular translation machinery components eukaryotic initiation factor (eIF)4E and eIF4G, but intact VP1 polymerase activity, VP3 dsRNA-binding activity, and the interaction between VP1 and VP3 are indispensable for both sense and antisense IBDV B MG activity. In addition, ribavirin, which inhibits IBDV replication, inhibits IBDV B MG activity in a dose-dependent manner. Collectively, the IBDV MG established in this study provides a powerful tool to investigate IBDV intracellular replication and transcription and virus‒host interactions and facilitates high-throughput screening for the identification of IBDV antivirals.
Insights
Researchers developed a novel infectious bursal disease virus (IBDV) minigenome (MG) system. This IBDV MG tool aids in studying viral replication and discovering new antiviral drugs.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Minigenomes (MGs) are crucial for studying RNA virus replication, transcription, and host interactions.
- An established MG system for infectious bursal disease virus (IBDV) was lacking, hindering research.
- IBDV is an important pathogen affecting poultry.
Purpose of the Study:
- To develop and characterize a functional minigenome system for IBDV.
- To utilize the IBDV MG for investigating viral replication mechanisms.
- To establish a platform for high-throughput screening of IBDV antivirals.
Main Methods:
- Constructed IBDV minigenomes by replacing coding sequences with reporter genes (Renilla luciferase or EGFP).
- Utilized the RNA polymerase I promoter for minigenome transcription.
- Investigated the role of viral proteins VP1 and VP3 in minigenome replication and translation.
- Assessed the impact of cellular translation factors (eIF4E, eIF4G) on minigenome activity.
- Evaluated the antiviral effect of ribavirin on the IBDV minigenome system.
Main Results:
- Successfully developed functional IBDV minigenomes (MGs) using reporter genes.
- IBDV B MG demonstrated higher activity compared to IBDV A MG.
- Sense IBDV B MG exhibited higher expression levels than antisense IBDV B MG.
- Minigenome activity depended on viral proteins VP1 and VP3, independent of cellular translation factors.
- Ribavirin inhibited IBDV B MG activity in a dose-dependent manner.
Conclusions:
- The established IBDV MG system is a valuable tool for studying IBDV replication and transcription.
- This system facilitates research on IBDV-host interactions.
- The IBDV MG provides a robust platform for high-throughput screening of potential antiviral compounds against IBDV.
