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Updated: May 14, 2026
Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay
Published on: February 22, 2018
[Cloning and functional research of Arp2/3-P40/ARPC1 subunit of Sf9 cells]
Shi-Li Han1, Jing-Fang Mu, Yong-Li Zhang
1Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Central China Normal University, Wuhan 430079, China. shanddonghanshili@126.com
Abstract:
The baculovirus-induced actin polymerization is mainly associated with the virus nucleocapsid protein P78/83, which is homologous with WASP proteins that can activate Arp2/3 complex and induce the actin polymerization. In order to explore the role of Arp2/3 complex in the baculovirus replication, the P40 subunit of Arp2/3 complex from Sf9 (Spodoptera frugiperda 9) cell line was cloned and characterized. Immunofluorescent microscopy assay indicated that P40 was recruited to the inner-side of nuclear membrane during virus infection, which was in accordance with nuclear F-actin distribution in virus-infected cells as documented in our previous research, suggesting P40 could be used to track Arp2/3 complex subcellular distribution changes during virus infection. In addition, co-immunoprecipitation assay demonstrated that P40 interacted with P78/83 only in virus-infected cells, suggesting that actin polymerization induced by P78/83-Arp2/3 complex during baculovirus infection was regulated by some unidentified virus factors.
Insights
Baculovirus infection triggers actin polymerization via nucleocapsid protein P78/83 interacting with Arp2/3 complex. This interaction, localized to the nuclear membrane, is regulated by viral factors, impacting baculovirus replication.
Area of Science:
- Cell Biology
- Virology
- Molecular Biology
Background:
- Baculovirus infection induces actin polymerization, primarily linked to the nucleocapsid protein P78/83.
- P78/83 is homologous to WASP proteins, known activators of the Arp2/3 complex, which drives actin polymerization.
Purpose of the Study:
- To investigate the role of the Arp2/3 complex in baculovirus replication.
- To characterize the P40 subunit of the Arp2/3 complex during baculovirus infection.
Main Methods:
- Cloning and characterization of the P40 subunit of Arp2/3 complex from Sf9 cells.
- Immunofluorescent microscopy to track P40 localization.
- Co-immunoprecipitation assays to study protein interactions.
Main Results:
- P40 localized to the inner nuclear membrane during baculovirus infection, correlating with nuclear F-actin distribution.
- P40 interacted with the viral protein P78/83 exclusively in infected cells.
Conclusions:
- The P40 subunit serves as a marker for Arp2/3 complex redistribution during baculovirus infection.
- Baculovirus-induced actin polymerization mediated by P78/83 and Arp2/3 complex is regulated by unidentified viral factors.
