Hepatitis B virus (HBV) genome transcription is highly dependent on liver-enriched, metabolic nuclear receptors (NRs). Among others, NR farnesoid X receptor alpha (FXRalpha) enhances HBV core promoter activity and pregenomic RNA synthesis. Interestingly, two food-withdrawal-induced FXRalpha modulators, peroxisome proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha) and deacetylase SIRT1, have been found to be associated with HBV genomes ex vivo. Whereas PGC-1alpha induction was shown to increase HBV replication, the effect of SIRT1 on HBV transcription remains unknown. Here, we showed that, in hepatocarcinoma-derived Huh-7 cells, combined activation of FXRalpha by GW4064 and SIRT1 by activator 3 increased HBV core promoter-controlled luciferase expression by 25-fold, compared with a 10-fold increase with GW4064 alone. Using cell lines differentially expressing FXRalpha in overexpression and silencing experiments, we demonstrated that SIRT1 activated the core promoter in an FXRalpha- and PGC-1alpha-dependent manner. Maximal activation (>150-fold) was observed in FXRalpha- and PGC-1alpha-overexpressing Huh-7 cells treated with FXRalpha and SIRT1 activators. Similarly, in cells transfected with full-length HBV genomes, maximal induction (3.5-fold) of core promoter-controlled synthesis of 3.5-kb RNA was observed in the same conditions of transfection and treatments. Thus, we identified a subnetwork of metabolic factors regulating HBV replication, strengthening the hypothesis that transcription of HBV and metabolic genes is similarly controlled.