Porcine circovirus type 2 infection promotes the SUMOylation of nucleophosmin-1 to facilitate the viral circular single-stranded DNA replication
Fig 5
PCV2 infection promotes Ubc9/TRIM24 signalings to enhance the SUMOylation of pNPM1.
(A, B) 1 MOI PCV2 or mock (the same volume of medium) infected PK-15 cells for 0 h, 6 h, 12 h, 18 h, and 24 h. (A) The expression levels of SAE1, Ubc9, TRIM24, TRIM62, and SENP3 were analyzed by western blot. (B) The expression levels of Ubc9 and SENP3 relative to β-actin were calculated. (C) The plasmids expressing TRIM24, TRIM62, and pNPM1 were transfected into 293T cells for 48 h, and the interactions of TRIM24 and TRIM62 with pNPM1 were detected by co-IP assays. (D) The plasmids expressing TRIM24, TRIM62, and pNPM1 were transfected into PK-15npm1-/- cells for 24 h, and then the cells were infected with 1 MOI PCV2 or mock for 12 h. The interactions of TRIM24 and TRIM62 with pNPM1 were detected by co-IP assays. (E) The specific siRNAs targeting TRIM24 or TRIM62 were transfected into PK-15 cells for 24 h, then the cells were infected with 1 MOI PCV2 for 12 h. The expression levels of TRIM24, TRIM62, pNPM1, and SUMOylated pNPM1 were detected by western blot and calculated. (F) The specific siRNA targeting Ubc9 was transfected into PK-15 cells for 24 h, then the cells were infected with 1 MOI PCV2 for 12 h. The interactions of TRIM24 with pNPM1 were detected by co-IP assays. **p < 0.01, *p < 0.05 versus the mock-infected cells at the same time point in (B), **p < 0.01 versus the siNC-transfected cells in (E), ##p < 0.01 versus the PCV2-infected cells at 18 h in (B).