Hepatitis B virus downregulates the human interferon-inducible MxA promoter through direct interaction of precore/core proteins.
The human MxA protein is an interferon (IFN)-inducible GTPase with proven antiviral activity against diverse viruses. IFN responsiveness is impaired in chronic hepatitis B virus (HBV) infection. Accordingly, initial experiments showed that, in contrast to parental HepG2 cells, when HepG2-derived 2.2.15 liver cells carrying the HBV genome were treated with IFN, they could not synthesize the MxA protein. Furthermore, MxA expression was reduced in HepG2 cells transiently transfected with the HBV genome. To assess whether HBV-encoded precore/core (preC/C) proteins interact with the IFN-signalling pathway, HepG2, Chang and HeLa cells were transfected with preC/C expression plasmids; the levels of signal transducers remained unaffected. Next, full-length and deletion mutants fused to the CAT reporter gene were tested to investigate whether MxA inhibition occurs at the promoter level. In co-transfection experiments, IFN-induced CAT activity was inhibited by preC/C expression in a dose-dependent manner. Analysis of deletion mutants showed that the region affected by the preC/C proteins comprises IFN-stimulated response elements 2 and 3, upstream of the putative start codon of the MxA promoter. In addition, HBV preC/C proteins interacted directly with the MxA promoter, as shown by electrophoretic mobility shift assays. These results demonstrate a mechanism that HBV probably uses to downregulate an element of the IFN-induced host antiviral responses, which accounts for the impairment observed in HBV-infected patients.
Fernández
Mario
M
Fundación para el Estudio de las Hepatitis Virales, C/Guzmán el Bueno 72, 28015 Madrid, Spain.
Quiroga
Juan Antonio
JA
Fundación para el Estudio de las Hepatitis Virales, C/Guzmán el Bueno 72, 28015 Madrid, Spain.
Carreño
Vicente
V
Fundación para el Estudio de las Hepatitis Virales, C/Guzmán el Bueno 72, 28015 Madrid, Spain.
D002460:Cell Line; D015536:Down-Regulation; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D006512:Hepatitis B Core Antigens / Q000235:genetics / Q000378:metabolism*; D006515:Hepatitis B virus / Q000472:pathogenicity*; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D064547:Myxovirus Resistance Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured
D006512:Hepatitis B Core Antigens; D016898:Interferon-alpha; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D011498:Protein Precursors; D019204:GTP-Binding Proteins