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Updated: Dec 27, 2025
Plaque Assay for Murine Norovirus
Published on: August 22, 2012
Effect of heat shock on murine norovirus replication in RAW264.7 cells
Chang Ma1, Xuliang Zhang1, Jinwei You1
1Department of Comparative Medicine, Jinling Hospital, Nanjing, PR China.
Abstract:
Murine norovirus (MNV), is a prevalent pathogen of laboratory mice closely related to human norovirus (HuNoV), a contagious pathogen known to cause gastroenteritis worldwide; however, the mechanism of norovirus replication remains poorly understood. Both heat shock protein 90 (Hsp90) and heat shock protein 70 (Hsp70) play an important role in viral genome replication and viral gene expression. In this study, we first found that heat stress exerted a positive effect on the replication of MNV in the murine macrophage RAW264.7 cell line. Inhibition of Hsp70 and Hsp90 by the specific inhibitors, KNK437 and 17-AGG, respectively showed that Hsp70 and Hsp90 enhanced MNV genome replication and virion production. In addition, we found that KNK437 and 17-AGG could decrease the level of IL-1β, IL-10, and TNF-α mRNA expression in MNV-infected cells. These data suggested that heat stress can positively regulate MNV replication, which advances our understanding of the molecular mechanism of MNV infection.
Insights
Heat stress enhances murine norovirus (MNV) replication by upregulating heat shock proteins (Hsp70 and Hsp90). Inhibiting these proteins reduced viral production and inflammatory cytokine mRNA levels in infected cells.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Murine norovirus (MNV) is a significant pathogen in laboratory mice, closely related to human norovirus (HuNoV).
- Understanding norovirus replication mechanisms is crucial due to the global impact of HuNoV gastroenteritis.
- Heat shock proteins (Hsp90 and Hsp70) are known to influence viral replication and gene expression.
Purpose of the Study:
- To investigate the effect of heat stress on MNV replication.
- To determine the role of Hsp70 and Hsp90 in MNV genome replication and virion production.
- To examine the impact of Hsp70 and Hsp90 inhibition on inflammatory cytokine expression during MNV infection.
Main Methods:
- Utilized the murine macrophage RAW264.7 cell line for MNV infection studies.
- Applied heat stress to infected cells to observe its effect on viral replication.
- Administered specific inhibitors (KNK437 for Hsp70, 17-AGG for Hsp90) to assess protein roles.
- Quantified viral genome replication and virion production.
- Measured mRNA expression levels of IL-1β, IL-10, and TNF-α.
Main Results:
- Heat stress was found to positively regulate MNV replication in RAW264.7 cells.
- Inhibition of Hsp70 and Hsp90 significantly enhanced MNV genome replication and virion production.
- Treatment with KNK437 and 17-AGG led to decreased levels of IL-1β, IL-10, and TNF-α mRNA in MNV-infected cells.
Conclusions:
- Heat stress positively influences MNV replication, likely through the action of Hsp70 and Hsp90.
- Hsp70 and Hsp90 play a critical role in promoting MNV genome replication and virion assembly.
- Modulation of Hsp70 and Hsp90 may impact the inflammatory response during MNV infection, offering insights into viral pathogenesis.
