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nsP3-FXR co-condensation enables alphavirus replication and reveals a targetable alphavirus vulnerability
Yifan Xie1, Jie Cao1, Xinya Yang2
1Life Sciences Institute, Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Zhejiang University, Hangzhou 310058, Zhejiang, China; Department of Infectious Diseases, Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Children and Adolescents' Health and Diseases, Hangzhou 310052, Zhejiang, China.
Abstract:
Alphaviruses are mosquito-borne RNA viruses that pose a significant threat to humans, with risks worsened by global warming. Non-structural protein 3 (nsP3) is critical for alphavirus infection, yet its precise roles remain poorly defined. Using Venezuelan equine encephalitis virus (VEEV) as a model, this study reveals that alphavirus unique domain (AUD) of nsP3 is essential for its condensation and viral replication. nsP3 forms gel-like condensates to bind to the second Tudor domain of Fragile X-related protein 1 (FXR1), forming replication supporting complexes. FXR1 bridges nsP3 and viral RNA within these condensates, which compartmentalize viral replication independent of protein interactions. Furthermore, UBAP2L was identified as a condensate-resident restriction factor against VEEV. Based on conserved AUD across alphaviruses, we developed a "21 R-AUD" antiviral strategy by fusing TRIM21's RING domain to VEEV's AUD. This approach induces proteasomal degradation of nsP3 condensates and inhibits replication of multiple alphaviruses, establishing conserved nsP3 condensation as a broad-spectrum antiviral target.
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