Increased phosphoenolpyruvate carboxykinase gene expression and steatosis during hepatitis C virus subgenome replication: role of nonstructural component 5A and CCAAT/enhancer-binding protein . Academic Article uri icon


  • Chronic hepatitis C virus (HCV) infection greatly increases the risk for type 2 diabetes and nonalcoholic steatohepatitis; however, the pathogenic mechanisms remain incompletely understood. Here we report gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK) transcription and associated transcription factors are dramatically up-regulated in Huh.8 cells, which stably express an HCV subgenome replicon. HCV increased activation of cAMP response element-binding protein (CREB), CCAAT/enhancer-binding protein (C/EBP), forkhead box protein O1 (FOXO1), and peroxisome proliferator-activated receptor coactivator 1 (PGC-1) and involved activation of the cAMP response element in the PEPCK promoter. Infection with dominant-negative CREB or C/EBP-shRNA significantly reduced or normalized PEPCK expression, with no change in PGC-1 or FOXO1 levels. Notably, expression of HCV nonstructural component NS5A in Huh7 or primary hepatocytes stimulated PEPCK gene expression and glucose output in HepG2 cells, whereas a deletion in NS5A reduced PEPCK expression and lowered cellular lipids but was without effect on insulin resistance, as demonstrated by the inability of insulin to stimulate mobilization of a pool of insulin-responsive vesicles to the plasma membrane. HCV-replicating cells demonstrated increases in cellular lipids with insulin resistance at the level of the insulin receptor, increased insulin receptor substrate 1 (Ser-312), and decreased Akt (Ser-473) activation in response to insulin. C/EBP-RNAi normalized lipogenic genes sterol regulatory element-binding protein-1c, peroxisome proliferator-activated receptor , and liver X receptor but was unable to reduce accumulation of triglycerides in Huh.8 cells or reverse the increase in ApoB expression, suggesting a role for increased lipid retention in steatotic hepatocytes. Collectively, these data reveal an important role of NS5A, C/EBP, and pCREB in promoting HCV-induced gluconeogenic gene expression and suggest that increased C/EBP and NS5A may be essential components leading to increased gluconeogenesis associated with HCV infection.

published proceedings

  • J Biol Chem

author list (cited authors)

  • Qadri, I., Choudhury, M., Rahman, S. M., Knotts, T. A., Janssen, R. C., Schaack, J., ... Friedman, J. E.

citation count

  • 20

complete list of authors

  • Qadri, Ishtiaq||Choudhury, Mahua||Rahman, Shaikh Mizanoor||Knotts, Trina A||Janssen, Rachel C||Schaack, Jerome||Iwahashi, Mieko||Puljak, Livia||Simon, Francis R||Kilic, Gordan||Fitz, J Gregory||Friedman, Jacob E

publication date

  • January 2012