Regulation of airway epithelial cell NF-kappa B-dependent gene expression by protein kinase C delta.

Abstract

Airway epithelial cells synthesize proinflammatory molecules such as IL-8, GM-CSF, RANTES, and ICAM-1, the expression of which is increased in the airways of patients with asthma. We investigated the regulation of these NF-kappa B-dependent genes by the novel protein kinase C (PKC) isoform PKC delta in 16HBE14o- human airway epithelial cells, focusing on IL-8 expression. Transient transfection with the constitutively active catalytic subunit of PKC delta (PKC delta-CAT), and treatment with bryostatin 1, an activator of PKC delta, each increased transcription from the IL-8 promoter, whereas overexpression of PKC epsilon had minor effects. Expression of a dominant negative PKC delta mutant (PKC delta-KR) or pretreatment of cells with rottlerin, a chemical PKC delta inhibitor, attenuated TNF-alpha- and phorbol ester-induced transcription from the IL-8 promoter. Bryostatin 1 treatment increased IL-8 protein abundance in primary airway epithelial cells. Selective activation of PKC delta by bryostatin also activated NF-kappa B, as evidenced by p65 RelA and p50 NF-kappa B1 binding to DNA, NF-kappa B trans-activation, and I kappa B degradation. The sufficiency of PKC delta to induce NF-kappa B nuclear translocation and binding to DNA was confirmed in a 16HBE14o- cell line inducibly expressing PKC delta-CAT under the tet-off system. Deletion of the NF-kappa B response element severely attenuated PKC delta-induced IL-8 promoter activity. Finally, PKC delta-CAT induced transcription from the GM-CSF, RANTES, and ICAM-1 promoters. Together these data suggest that PKC delta plays a key role in the regulation of airway epithelial cell NF-kappa B-dependent gene expression.

• Authors: Kristen Page, Jing Li, Limei Zhou, Svetlana Iasvovskaia, Kevin C Corbit, Jae-Won Soh, I Bernard Weinstein, Allan R Brasier, Anning Lin, Marc B Hershenson, Svetlana Iasvoyskaia
• Journal: Journal of immunology (Baltimore, Md. : 1950) (J Immunol) Vol. 170 Issue 11 Pg. 5681-9 (Jun 01 2003) ISSN: 0022-1767 [Print] United States
• PMID: 12759450 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Chemokine CCL5
• DNA-Binding Proteins
• Interleukin-8
• Isoenzymes
• NF-kappa B
• Trans-Activators
• Intercellular Adhesion Molecule-1
• Granulocyte-Macrophage Colony-Stimulating Factor
• Protein Serine-Threonine Kinases
• CHUK protein, human
• I-kappa B Kinase
• IKBKB protein, human
• IKBKE protein, human
• PRKCD protein, human
• Protein Kinase C
• Protein Kinase C-delta

Topics

• Cell Line
• Cells, Cultured
• Chemokine CCL5 (genetics, physiology)
• DNA-Binding Proteins (metabolism, physiology)
• Enzyme Activation (genetics)
• Gene Expression Regulation (physiology)
• Granulocyte-Macrophage Colony-Stimulating Factor (genetics, physiology)
• Humans
• I-kappa B Kinase
• Intercellular Adhesion Molecule-1 (genetics, physiology)
• Interleukin-8 (biosynthesis, genetics)
• Isoenzymes (biosynthesis, physiology)
• NF-kappa B (physiology)
• Promoter Regions, Genetic (physiology)
• Protein Kinase C (biosynthesis, physiology)
• Protein Kinase C-delta
• Protein Serine-Threonine Kinases (metabolism, physiology)
• Respiratory Mucosa (cytology, enzymology, metabolism, pathology)
• Trans-Activators (metabolism, physiology)
• Transcription, Genetic