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Aim: To explore the effects of HSP90 on rotavirus (RV) replication. Materials & methods: The interaction between NSP2 and HSP90 was analyzed by immunoprecipitation assay, and co-localization confirmed by an immunofluorescence assay. The viral titer was measured by immunofluorescence assay and viral protein expression investigated by western blotting. Results: The interaction between NSP2 and HSP90 was confirmed. Knockdown of HSP90 using siRNA resulted in a decrease in NSP2 expression and virus titer. HSP90 inhibitor 17-AAG significantly decreased NSP2 and NSP5 expression, as well as the virus titer, while proteasome inhibitor MG132 restored the protein amount and the viral titer. Conclusion: HSP90 is involved in rotavirus replication, and inhibition of HSP90 promotes the degradation of NSP2 through the proteasome pathway. Rotavirus is a type of virus that causes severe diarrhea, especially in children. The virus infects the cell in our bodies. These cells have proteins which carry out important functions and can help the growth of the virus. One of these proteins, called HSP90, interacts with a protein of the virus, called NSP2. After infection, these proteins move to the same place within the cell. Decreasing the level of HSP90 also decreases the level of NSP2, slowing the growth of the virus. Understanding how HSP90 and NSP2 work will help to develop new drugs to treat rotavirus infection. Article highlights Background Heat shock protein 90 (HSP90) appears to play an essential role in viral replication, but little is known regarding the role of HSP90 in rotavirus (RV) infection. Methods RV laboratory strain SA11 was used in this study. Virus titers were measured by Fluorescent formation units assay and viral protein expression checked by WB. The interaction of HSP90 and NSP2 was confirmed by immunoprecipitation (IP) and confocal laser microscope. siRNA used to knockdown the expression of HSP90 and HSP90 activity was inhibited. Results The HSP90 inhibition resulted in decreased RV replication. Knockdown of HSP90 by siRNA decreased RV protein levels and virus replication. HSP90 interacted with virus protein NSP2. Inhibition of the proteasome pathway decreased viral protein degradation. Conclusion HSP90 was used by RV to facilitate virus replication by chaperoning viral proteins and preventing their degradation via the proteasome. [ABSTRACT FROM AUTHOR]

Aim: The preparation of a VSVΔG*-S pseudovirus carrying the spike (S) protein of SARS-CoV-2 and its preliminary application. Materials & methods: 293T cells were transfected with pCAGGS-SARS-2-S and infected with VSVΔG viruses. The VSVΔG*-S pseudovirus was collected by centrifugation. The pseudovirus was used to investigate the neutralization ability of serum from SARS-CoV-2 patients and the ability of peptides to inhibit the virus. Results: A VSVΔG*-S pseudovirus was successfully prepared. Patient serum and peptides could neutralize the pseudovirus infection as determined by measuring the expression of GFP. Conclusion: This pseudovirus system can be used to screen polypeptides and fusion inhibitors and measure neutralizing activity, which will benefit the study of SARS-CoV-2. The virus SARS-CoV-2 is the cause of COVID-19 disease. This virus is harmful and easily spreads between people, so when being worked with in a laboratory, it must be under high-security conditions. Laboratories with this level of safety are not common, so this reduces the ability to study SARS-CoV-2. In this study, we engineered a modified version of the virus, called a pseudovirus, that can be worked with in a lower security laboratory. [ABSTRACT FROM AUTHOR]