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Published on: January 13, 2022
Deletion of AcMNPV ac146 eliminates the production of budded virus
Virginia L Dickison1, Leslie G Willis, Nadia R Sokal
1Irving K. Barber School of Arts and Sciences, Unit 2 Biology, University of British Columbia, Kelowna, B.C. V1V 1V7, Canada.
Abstract:
Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac146 is a highly conserved gene in the Alpha- and Betabaculovirus genera that has an unknown function. Northern blot analysis and transcript mapping showed that ac146 is transcribed at late times post infection as a 1.2 kb mRNA. To determine the role of ac146 in the baculovirus life cycle ac146 knock out viruses were constructed. Transfection and plaque assays showed that all the ac146 deletions produced a single cell phenotype indicating that no infectious budded virus (BV) was produced, however occlusion bodies were formed. The lack of BV production was confirmed by viral titration utilizing both qPCR and TCID₅₀. Analysis of BV and occlusion derived virus (ODV) revealed that AC146 is associated with both forms of the virus and is modified specifically in ODV. This study therefore demonstrates that AC146 is a late virion associated protein and is essential for the viral life cycle.
Insights
The Autographa californica multiple nucleopolyhedrovirus ac146 gene is essential for producing infectious budded virus (BV). Deleting ac146 prevents BV formation, highlighting its critical role in the baculovirus life cycle.
Area of Science:
- * Virology
- * Molecular Biology
- * Insect Pathology
Background:
- * The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac146 gene is highly conserved in baculoviruses.
- * The function of ac146 has remained unknown despite its conservation.
Purpose of the Study:
- * To elucidate the function of the ac146 gene in the baculovirus life cycle.
- * To determine the role of AC146 protein during viral infection and replication.
Main Methods:
- * Northern blot analysis and transcript mapping to characterize ac146 expression.
- * Construction and analysis of ac146 knockout viruses.
- * Viral titration using qPCR and TCID₅₀ assays.
Main Results:
- * ac146 is transcribed as a 1.2 kb mRNA at late times post-infection.
- * ac146 knockout viruses failed to produce infectious budded virus (BV) but formed occlusion bodies.
- * AC146 protein is associated with both budded virus (BV) and occlusion-derived virus (ODV), with specific modification in ODV.
Conclusions:
- * AC146 is a late, virion-associated protein.
- * AC146 is essential for the production of infectious budded virus (BV) and the overall baculovirus life cycle.
