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Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay
Published on: February 22, 2018
Analysis of host specificity of two closely related baculoviruses in permissive and nonpermissive cell lines
Md Masmudur Rahman1, Karumathil P Gopinathan
1Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India.
Abstract:
The baculoviruses Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) share about 90% identity at the genomic level but they have non-overlapping host range and show a high degree of host specificity. We have demonstrated here that AcMNPV undergoes DNA replication and early gene expression in Bombyx-derived BmN cells but fails to show very late gene expression or produce budded virion (BV) particles. Coinfection with BmNPV supported BV production from AcMNPV in BmN cells at low levels but not very late gene expression or polyhedral inclusion body formation. BV production and very late gene expression from BmNPV, on the contrary, were adversely affected in coinfections. In Spodoptera frugiperda-derived Sf21 cell lines, BmNPV DNA replication, BV production, and very late gene expression took place only when coinfected with AcMNPV. BmNPV exerted a less profound effect on AcMNPV multiplication and very late gene expression in permissive host cell lines. AcMNPV shuts down cellular and viral protein synthesis completely when infected alone or coinfected with BmNPV in BmN cells, whereas BmNPV infection did not affect cellular and viral protein synthesis in Sf21 cells. Overall, AcMNPV showed a more dominant effect by complementing the multiplication of BmNPV in nonpermissive host cells while inhibiting it in BmN cells.
Insights
Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) interactions were studied. AcMNPV complements BmNPV replication in non-permissive cells, while inhibiting BmNPV in permissive cells.
Area of Science:
- Virology
- Molecular Biology
- Insect Pathology
Background:
- Baculoviruses, including Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV), exhibit high genomic similarity but distinct host ranges.
- Understanding viral interactions and host specificity is crucial for baculovirus applications.
Purpose of the Study:
- To investigate the interactive effects of BmNPV and AcMNPV coinfection on viral replication and gene expression in different insect cell lines.
- To elucidate the mechanisms underlying baculovirus host specificity and cross-interference.
Main Methods:
- Coinfection experiments using BmN (Bombyx mori-derived) and Sf21 (Spodoptera frugiperda-derived) cell lines.
- Analysis of viral DNA replication, early and very late gene expression, and budded virion (BV) production.
Main Results:
- AcMNPV supported BmNPV replication and BV production in Sf21 cells, but inhibited BmNPV in BmN cells.
- BmNPV adversely affected AcMNPV BV production and very late gene expression in BmN cells.
- AcMNPV demonstrated a dominant effect, complementing BmNPV in non-permissive cells while inhibiting it in permissive cells, and shut down protein synthesis in BmN cells.
Conclusions:
- AcMNPV plays a dominant role in modulating the replication of both viruses, influencing host range and viral interactions.
- The findings highlight complex regulatory mechanisms governing baculovirus replication and host specificity.
- Differential effects on protein synthesis indicate distinct viral strategies impacting cellular processes.
