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Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis
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The Function behind the Relation between Lipid Metabolism and Vimentin on H9N2 Subtype AIV Replication
Anran Lu1,2, Jing Yang1,3, Xiangyu Huang2
1Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Viruses
|August 26, 2022
Summary
Avian influenza virus H9N2 infection increases cholesterol and decreases HMGCR phosphorylation. Vimentin impacts lipid regulation, suggesting lipid rafts and cholesterol are key targets for antiviral strategies against H9N2.
Area of Science:
- Virology
- Cell Biology
- Biochemistry
Background:
- Avian influenza virus (AIV) subtype H9N2 threatens the poultry industry.
- Vimentin's role in lipid metabolism during viral infection is not fully understood.
- Understanding lipid metabolism's impact on H9N2 AIV replication is crucial.
Purpose of the Study:
- To investigate the function of lipid metabolism and vimentin in H9N2 AIV replication.
- To analyze cholesterol levels and HMGCR phosphorylation in vimentin knockout and HeLa cells with/without H9N2 AIV infection.
Main Methods:
- Comparative analysis of cholesterol content and HMGCR phosphorylation in vimentin KO and HeLa cells.
- Assessment of viral replication inhibition using cholesterol inhibitor lovastatin.
- Investigation of adenylate activated protein kinase (AMPK) activity and phosphorylation.
- Evaluation of viral hemagglutinin (HA) gene expression after MβCD treatment and cholesterol replenishment.
Main Results:
- H9N2 AIV infection increased cholesterol and reduced HMGCR phosphorylation in both cell types.
- Viral replication was inhibited by lovastatin.
- AMPK activity was reduced in infected cells, with lower phosphorylation in KO cells.
- Disrupting lipid rafts decreased viral HA gene levels in HeLa cells but increased them in KO cells.
Conclusions:
- Lipid rafts and cholesterol levels are critical for H9N2 AIV infection in human cells.
- Vimentin plays a significant role in regulating lipid effects on viral replication.
- Targeting lipid metabolism and vimentin offers potential antiviral strategies against influenza viruses.
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