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Originally published In Press as doi:10.1074/jbc.M005227200 on July 28, 2000

J. Biol. Chem., Vol. 275, Issue 41, 32250-32259, October 13, 2000
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Phorbol Ester-induced Expression of Airway Squamous Cell Differentiation Marker, SPRR1B, Is Regulated by Protein Kinase Cdelta /Ras/MEKK1/MKK1-dependent/AP-1 Signal Transduction Pathway*

Hue VuongDagger , Tricia PattersonDagger , Paul Shapiro§, Dhananjaya V. Kalvakolanu, Reen Wu||, Wei-Ya Ma**, Zigang Dong**, Steven R. KleebergerDagger , and Sekhar P. M. ReddyDagger Dagger Dagger

From the Dagger  Department of Environmental Health Sciences, The Johns Hopkins University School of Public Health, Baltimore, Maryland 21205, § University of Maryland School of Pharmacy, Baltimore, Maryland 21201,  Greenbaum Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland 21201, the || Department of Internal Medicine, School of Medicine, University of California, Davis, California 95616, and ** The Hormel Institute, University of Minnesota, Minneapolis, Minnesota 55912

The transcriptional induction of SPRR1B by phorbol 12-myristate 13-acetate (PMA) is mainly mediated by the first -152-base pair 5'-flanking region containing two functional AP-1 sites. In this study, we have analyzed the signaling pathways that mediate the induction in tracheobronchial epithelial cells. PKC inhibitor ablated PMA-stimulated expression of endogenous SPRR1B and reporter gene expression driven by SPRR1B promoter. PKC activator promoted the transcription. The dominant negative protein kinase Cdelta (dn-PKCdelta ) and rottlerin (PKCdelta inhibitor) completely suppressed PMA-stimulated promoter activity. dn-Ras or dn-MEKK1 inhibited PMA-stimulated promoter activity, while their corresponding constitutively active mutants augmented it. dn-c-Raf-1 did not have any effect on reporter gene expression. Since MEKK1 activates multiple parallel pathways, we examined involvement of JNK/SAPK, p38, and MKK1 in promoter regulation. Co-expression of the dominant negative forms of MKK4, MKK7, JNK/SAPK, MKK3, MKK6, or p38alpha did not suppress PMA-stimulated reporter gene expression. However, MKK1 inhibitors UO126 and PD98095 suppressed gene expression. Consistent with this, expression of dn-MKK1 strongly suppressed PMA-stimulated promoter activity, while the constitutively active MKK1 augmented it. However, MKK1-mediated induction of SPRR1B probably does not depend on extracellular signal-regulated kinases 1 and 2, suggesting the requirement of another kinase(s). dn-c-Jun mutants abolished PMA-stimulated expression supporting an important role for AP-1 proteins in SPRR1B expression. Together, these results suggest that a PKCdelta /Ras/MEKK1/MKK1-dependent/AP-1 pathway regulates the PMA-inducible expression of the SPRR1B in tracheobronchial epithelial cells.

* This work was supported in part by National Institutes of Health Grants HL-58122 (to S. R.), ES-06230 (to R. W.), CA-71401 and CA-78282 (to D. K.), and HL-57142 (to S. K.) and a grant from the Maryland Thoracic Society (to S. R.). Core facilities at the Johns Hopkins Urban Environmental Health Center were supported by NIEHS, National Institutes of Health, Grant ES03819.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Dagger To whom all correspondence should be addressed: Dept. of Environmental Health Sciences, The Johns Hopkins University, Division of Physiology, Rm. W7006, 615 N. Wolfe St., Baltimore, MD 21205. Tel.: 410-614-5442; Fax: 410-955-0299; E-mail: sreddy@jhsph.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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