Fig. 1: PWWP3A undergoes polyubiquitination and degradation in response to RNA virus infection.

The human THP-1 and HEK293 cells (a), as well as primary V5-Pwwp3a knock-in MEFs (b) were infected with sendai virus (SeV) (a, b, c), vesicular stomatitis virus (VSV) and herpes simplex virus type 1(HSV-1) (b) at the indicated time points for the immunoblotting analysis. d RT-qPCR analysis of PWWP3A (orange) in human THP-1 and HEK293 cells (n = 3 biological replicates per group). Primary V5-Pwwp3a knock-in MEFs (e) or HEK293 cells (f) were infected with SeV at the indicated time points, with or without cycloheximide (CHX) treatment (50 μM) for the immunoblotting analysis. g HEK293 cells infected with SeV at the indicated time points, with or without MG132 treatment (25 μM) for the immunoblotting analysis. h HEK293 cells were infected with SeV at the indicated time points for the co-immunoprecipitation (Co-IP) and immunoblot analysis. PWWP3A-Ubi (K48-linked), the lysine 48-linked ubiquitination chains enriched in PWWP3A proteins. i Protein-protein interaction (PPI) map for PWWP3A and E3 ubiquitin ligases. j The control (con.) and PJA2-knockout (KO) HEK293 cells were uninfected or infected with SeV for 8 h for the immunoblotting analysis. HEK293 cells were transfected with the indicated plasmids (k) or infected with SeV at the indicated time points (l) for the co-IP assays and immunoblotting analysis. m HEK293 cells were transfected with the indicated plasmids for the denature-immunoprecipitation (Denature-IP) assays. K48O/K63O, all the lysine residues in ubiquitin were mutated to arginine except K48 (K48O) or K63 (K63O). n The control and PJA2-KO HEK293 cells were infected with SeV at the indicated time point for the Co-IP assays and immunoblotting analysis. Graphs show mean ± SD, p values are from one-way ANOVA (d). At least three independent experiments were performed with similar results. Source data are provided as a Source Data file.