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-INDUCED C/EBP ISOFORMS
1 Pulmonary Center, Boston University School of Medicine, Boston, MA, USA
2 Pulmonary Center, Boston University School of Medicine, Boston, MA, USA; Pulmonary Medicine, Boston VA Medical Center, Boston, MA, USA
* To whom correspondence should be addressed. E-mail: pkuang{at}lung.bumc.bu.edu.
We previously showed that interleukin-1
(IL-1) decreased elastin gene transcription through activation of the NF-
B subunit p65 in neonatal rat lung fibroblasts. The present study was undertaken to further explore the molecular mechanisms responsible for the inhibitory effect of IL-1 on elastin gene transcription. We found that cycloheximide blocked IL-1-induced down-regulation of elastin mRNA but did not inhibit IL-1-induced translocation of p65 into the nucleus. IL-1 treatment increased CCAAT/enhancer-binding protein- (C/EBP) mRNA and protein levels including LAP (liver-enriched activating protein) and LIP (liver-enriched inhibitory protein) that was cycloheximide sensitive. C/EBP isoforms bound a GCAAT containing sequence in the proximal elastin promoter as determined by electrophoretic gel shift studies and confirmed by using specific C/EBP antibodies and by competition studies with oligonucleotides. Transient transfection of LIP expression vectors strongly decreased the transcriptional activity of the co-transfected elastin promoter and decreased levels of endogenous elastin mRNA. We demonstrated IL-1 induced down-regulation of elastin mRNA was dependent on NF-B activation and C/EBP expression. These results indicate that IL-1 treatment activates NF-B that subsequently induces LIP expression and inhibition of elastin gene transcription.
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