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Am J Physiol Lung Cell Mol Physiol 282: L215-L225, 2002; doi:10.1152/ajplung.00125.2001
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Vol. 282, Issue 2, L215-L225, February 2002

JNK1 and AP-1 regulate PMA-inducible squamous differentiation marker expression in Clara-like H441 cells

Hue Vuong1, Tricia Patterson1, Pavan Adiseshaiah1, Paul Shapiro2, Dhananjaya V. Kalvakolanu3, and Sekhar P. M. Reddy1

1 Department of Environmental Health Sciences, The Johns Hopkins University School of Public Health, Baltimore 21205; 2 University of Maryland School of Pharmacy, Baltimore 21201; and 3 Greenbaum Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland 21201

Exposure of distal bronchiolar region to various toxicants and pollutants suppresses Clara cell differentiation marker expression and greatly enhances the induction of squamous cell differentiation (SCD). Here, we demonstrate for the first time phorbol 13-myristate 12-acetate (PMA)-inducible expression of SCD markers, SPRRs, in Clara-like H441 cells. The transcriptional stimulation of human SPRR1B expression is mainly mediated by a -150- to -84-bp region that harbors two critical activator protein (AP)-1 sites. In unstimulated cells, the -150- to -84-bp region is weakly bound by AP-1 proteins, mainly JunD and Fra1. However, PMA prominently induced the binding of JunB and Fra1. Consistent with this, overexpression of wild-type Jun proteins upregulated the SPRR1B promoter activity. Conversely, a c-jun mutant suppressed both basal and PMA-inducible reporter gene expression. Intriguingly, overexpression of fra2 suppressed PMA-inducible reporter activity, whereas fra1 significantly enhanced basal level activity, indicating an opposing role for these proteins in SPRR1B expression in a manner similar to that observed in proximal tracheobronchial epithelial cells (BEAS-2B clone S6). Interestingly, unlike in S6 cells, a catalytically inactive c-Jun NH2-terminal kinase (JNK) 1 mutant significantly reduced the PMA-inducible SPRR1B promoter activity in H441 cells. Thus either temporal expression and/or spatial activation of AP-1 proteins by JNK1 might contribute to the induction of SCD in Clara cells.

small proline rich proteins; airway epithelium; transcriptional regulation; mitogen-activated protein kinases; Jun/Fos; c-Jun NH2-terminal kinase 1; activator protein 1


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