Regulation of airway epithelial cell NF-kappa B-dependent gene expression by protein kinase C delta.
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.
Page
Kristen
K
Department of Pediatrics, University of Chicago, Chicago, IL 60637, USA.
Li
Jing
J
Zhou
Limei
L
Iasvovskaia
Svetlana
S
Corbit
Kevin C
KC
Soh
Jae-Won
JW
Weinstein
I Bernard
IB
Brasier
Allan R
AR
Lin
Anning
A
Hershenson
Marc B
MB
Iasvoyskaia
Svetlana
S
D002460:Cell Line; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology; D004789:Enzyme Activation / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000502:physiology; D006801:Humans; D051550:I-kappa B Kinase; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000502:physiology; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D007527:Isoenzymes / Q000096:biosynthesis / Q000502:physiology; D016328:NF-kappa B / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000502:physiology; D011493:Protein Kinase C / Q000096:biosynthesis / Q000502:physiology*; D051745:Protein Kinase C-delta; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism / Q000502:physiology; D020545:Respiratory Mucosa / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism* / Q000473:pathology; D015534:Trans-Activators / Q000378:metabolism / Q000502:physiology; D014158:Transcription, Genetic
D018946:Chemokine CCL5; D004268:DNA-Binding Proteins; D016209:Interleukin-8; D007527:Isoenzymes; D016328:NF-kappa B; D015534:Trans-Activators; D018799:Intercellular Adhesion Molecule-1; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D017346:Protein Serine-Threonine Kinases; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human; C497258:PRKCD protein, human; D011493:Protein Kinase C; D051745:Protein Kinase C-delta