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Fig. 5 | Retrovirology

Fig. 5

From: Trim28 acts as restriction factor of prototype foamy virus replication by modulating H3K9me3 marks and destabilizing the viral transactivator Tas

Fig. 5

Trim28 decreases the expression of Tas protein and inhibits PFV replication through ubiquitin–proteasome pathway. a Trim28 down regulates the expression of Tas protein and the inhibition was reversed by MG132. HA-Tas combined with Flag-Trim28 or pCMV-Flag (as control) were transfected into HEK293T cells; 8 h before harvest, the cells were treated with DMSO or MG132 (5 Î¼M). Western blot was performed to assess Tas protein levels; b Trim28 promotes Tas polyubiquitination. HA-tagged ubiquitin (HA-Ub), Myc-Tas combined with Flag-Trim28 were transfected into HEK293T cells, pCMV-Flag used as negative control. c MG132 rescues the suppression of PFV produced by overexpressed Trim28, and deletion of RBCC domain abrogates its inhibitory effect. HT1080 cells were transfected with Flag-Trim28, HA-Trim28-â–³RBCC or pCMV-Flag (as control) for 24 h and challenged with PFV (MOI = 0.1) for another 48 h. 8 h before harvest, one group of cells transfected with Flag-Trim28 were treated with MG132 (5 μM, another groups were treated with DMSO). d Deletion of RBCC domain of Trim28 cannot cause trigger the ubiquitination of Tas. In cellular ubiquitination assay, HEK293T cells were transfected with HA-tagged ubiquitin (HA-Ub), Myc-Tas combined with Flag-Trim28 or Flag-Trim28-â–³RBCC, pCMV-Flag used as negative control

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Retrovirology

ISSN: 1742-4690

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