Phosphoproteomic Profiling Reveals Epstein-Barr Virus Protein Kinase Integration of DNA Damage Response and Mitotic Signaling
Fig 8
Regulation of known APC/C substrates by BGLF4.
(A) Fold change in protein level of APC/C substrates as measured by MS. See also S2 Table. (B) Western blot validation of BGLF4 mediated up-regulation of individual APC/C substrates in Dox inducible Akata (EBV+) cells expressing wild-type BGLF4, but not SUMO binding-deficient and kinase-dead mutants (BGLF4mSIM-N and BGLF4KD). Immunoblot analysis of the same cell lysates from Fig 2C using antibodies as indicated. (C) EBV replication induces the accumulation of TOP2A and Aurora B. Western blot analysis of cell extracts from Akata-BX1 (EBV+) treated as indicated using anti-Aurora B, anti-TOP2A and anti-β-actin antibodies as indicated. (D) and (E) SAC inhibition suppresses production of extracellular EBV virus. Supernatant virion DNA (D) and cell associated viral DNA (E) from Akata (EBV+) cells treated as indicated was determined by PCR. The experiments were carried out in three biological replicates with similar results and the representative results are presented. Relative PCR product intensity was quantified by ImageJ software.