In rat aortic smooth muscle cells (RASMC), interferon (IFN)-γ enhanced nitrite accumulation and type II nitric oxide synthase (iNOS) protein expression induced by interleukin (IL)-1β. IFN-γ alone had no effect on nitrite accumulation or iNOS protein. IL-1β, but not IFN-γ, induced nuclear factor (NF)-κB and CCAAT box/enhancer binding protein (C/EBP) nuclear binding. Conversely, IFN-γ, but not IL-1β, induced signal transducer and activator of transcription (STAT) 1 and interferon regulatory factor (IRF)-1 binding. In a -1.4-kb rat iNOS promoter segment, deletion of an IFN-γ-activated site (GAS) increased IL-1β-induced activity but inhibited IFN-γ-enhanced activity, suggesting a two-way effect of the GAS site on iNOS induction: enhancing induction through STAT1 activation and inhibiting induction through a non-IFN-γ-mediated mechanism. Deletion of both an IRF and a C/EBP site reduced the IL-1β-induced and the IFN-γ-enhanced activities. However, IRF site mutations decreased the IFN-γ-enhanced activity without affecting the IL-1β-induced activity. Insertion of two IRF sites increased the IFN-γ-enhanced, but not the IL-1β-induced, activity. Mutations of a reverse NF-κB site did not significantly change IFN-γ-enhanced activity. We conclude that in RASMC, NF-κB and C/EBP mediate the IL-1β-induced iNOS expression, whereas IRF-1 and STAT1 mediate the IFN-γ-enhanced iNOS induction.
- CCAAT box/enhancer binding protein
- Interferon regulatory factor-1
- Interferon-γ activation site
- Nuclear factor-κB
- Signal transducer and activator of transcription 1
ASJC Scopus subject areas
- Cell Biology