Host phosphatidic acid phosphatase lipin1 is rate limiting for functional hepatitis C virus replicase complex formation

PLoS Pathog. 2018 Sep 18;14(9):e1007284. doi: 10.1371/journal.ppat.1007284. eCollection 2018 Sep.

Abstract

Hepatitis C virus (HCV) infection constitutes a significant health burden worldwide, because it is a major etiologic agent of chronic liver disease, cirrhosis and hepatocellular carcinoma. HCV replication cycle is closely tied to lipid metabolism and infection by this virus causes profound changes in host lipid homeostasis. We focused our attention on a phosphatidate phosphate (PAP) enzyme family (the lipin family), which mediate the conversion of phosphatidate to diacylglycerol in the cytoplasm, playing a key role in triglyceride biosynthesis and in phospholipid homeostasis. Lipins may also translocate to the nucleus to act as transcriptional regulators of genes involved in lipid metabolism. The best-characterized member of this family is lipin1, which cooperates with lipin2 to maintain glycerophospholipid homeostasis in the liver. Lipin1-deficient cell lines were generated by RNAi to study the role of this protein in different steps of HCV replication cycle. Using surrogate models that recapitulate different aspects of HCV infection, we concluded that lipin1 is rate limiting for the generation of functional replicase complexes, in a step downstream primary translation that leads to early HCV RNA replication. Infection studies in lipin1-deficient cells overexpressing wild type or phosphatase-defective lipin1 proteins suggest that lipin1 phosphatase activity is required to support HCV infection. Finally, ultrastructural and biochemical analyses in replication-independent models suggest that lipin1 may facilitate the generation of the membranous compartment that contains functional HCV replicase complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Hepacivirus / genetics
  • Hepacivirus / pathogenicity*
  • Hepacivirus / physiology*
  • Hepatitis C / etiology
  • Hepatitis C / metabolism*
  • Hepatitis C / virology*
  • Host-Pathogen Interactions
  • Humans
  • Lipid Metabolism
  • Phosphatidate Phosphatase / antagonists & inhibitors
  • Phosphatidate Phosphatase / genetics
  • Phosphatidate Phosphatase / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Viral / genetics
  • RNA, Viral / metabolism
  • RNA-Dependent RNA Polymerase / metabolism*
  • Virus Replication

Substances

  • RNA, Messenger
  • RNA, Small Interfering
  • RNA, Viral
  • RNA-Dependent RNA Polymerase
  • LPIN1 protein, human
  • Phosphatidate Phosphatase

Grants and funding

This study was funded by Ministerio de Economía y Competitividad grants: SAF2010-19270, SAF2013-44282-R, SA2014-52400-R. VC was supported by an FPI fellowship from Ministerio de Economía y Competitividad (BES2015-07492). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.