F1Fo ATP synthase is present in all organisms and is predominantly located on the inner membrane of mitochondria in eukaryotic cells. The present study demonstrated that ATP synthase and electron transport chain complexes were ectopically expressed on the surface of breast cancer cells and could serve as a potent anticancer target. We investigated the anticancer effects of the ATP synthase inhibitor citreoviridin on breast cancer cells through proteomic approaches and revealed that differentially expressed proteins in cell cycle regulation and in the unfolded protein response were functionally enriched. We showed that citreoviridin triggered PERK-mediated eIF2α phosphorylation, which in turn attenuated general protein synthesis and led to cell cycle arrest in the G0/G1 phase. We further showed that the combination of citreoviridin and the 26S proteasome inhibitor bortezomib could improve the anticancer activity by enhancing ER stress, by ameliorating citreoviridin-caused cyclin D3 compensation, and by contributing to CDK1 deactivation and PCNA downregulation. More interestingly, the combined treatment triggered lethality through unusual non-apoptotic caspase- and autophagy-independent cell death with a cytoplasmic vacuolization phenotype. The results imply that by boosting ER stress, the combination of ATP synthase inhibitor citreoviridin and 26S proteasome inhibitor bortezomib could potentially be an effective therapeutic strategy against breast cancer.

Transcription of the human polyomavirus (JCV) genome is regulated by host cell proteins and the viral early protein, T antigen. A region called the lytic control element (LCE), located within the enhancer of JCV, is important for transcription of JCV early and late promoters. Earlier studies have led to the identification of two single-stranded DNA-binding proteins, YB-1 and Pur alpha, with the ability to interact with nucleotides on the early and late strands of LCE, respectively. Of particular interest is the notion that the unique interplay between these two cellular proteins and JCVT antigen determines their binding activities with the LCE. In this study, we employed a series of cotransfection experiments to evaluate the levels of transcription from JCV early and late promoters in the presence of YB-1, Pur alpha, and T antigen. Results from these studies indicated that Pur alpha stimulates JCV early and has little effect on the late promoter. Moreover, T antigen was able to decrease the induced level of early gene transcription by Pur alpha. On the other hand, the extent of transactivation of the viral late promoter by T antigen was reduced upon overexpression of Pur alpha in the transfected cells. These observations suggest that Pur alpha and T antigen exert an antagonistic effect on each other's regulatory action upon their responsive promoters. Of particular interest was the observation that YB-1 liberated T-antigen-induced late promoter activity from repression imposed by overexpression of pur alpha. Under similar conditions, overexpression of YB-1 showed no effect on the transcriptional activity of the early promoter in cells transfected with T-antigen- and Pur alpha-producing plasmids. On the basis of the data presented here and the previous binding results, a model is proposed which describes the potential role of Pur alpha, YB-1, and T antigen in differential expression of the viral genome during the lytic cycle.