p11 expression in human bronchial epithelial cells is increased by nitric oxide in a cGMP-dependent pathway involving protein kinase G activation.
The effect of nitric oxide on p11 expression was studied in an immortalized human bronchial epithelial cell line (BEAS-2B cells). Three nitric oxide donors were used: spermine NONOate (SP), (+/-)-S-nitroso-N-acetylpenicillamine (SNAP), and S-nitrosoglutathione (SNOG). All three nitric oxide donors had similar effects resulting in dose-dependent and time-dependent accumulation of p11 protein and an increase of steady-state p11 mRNA. Studies using a reporter gene containing the region from -1499 to +89 of the p11 promoter demonstrated an increase in transcriptional activity after stimulation with NO donors for 4 h. These effects were abolished at the promoter and protein level using protein kinase G inhibitors (KT5823 and R(p)-8-pCPT-cGMPS). Incubation of transfected cells with a cell permeable cGMP analogue (8-Br-cGMP) resulted in a dose-related increase of promoter activity. An electrophoretic mobility shift assay of nuclear proteins extracted from BEAS-2B cells identified an AP-1 site located at -82 to -77 of the p11 promoter region as an NO- and cGMP- dependent response element. These data were confirmed using a c-jun dominant negative mutant vector and a c-jun expression plasmid. Therefore, we conclude that nitric oxide-induced p11 expression in human bronchial epithelial cells is mediated at least in part through increased binding of activator protein one to the p11 promoter.
Pawliczak
R
R
Critical Care Medicine Department, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland 20892, USA.
Cowan
M J
MJ
Huang
X
X
Nanavaty
U B
UB
Alsaaty
S
S
Logun
C
C
Shelhamer
J H
JH
D000470:Alkaloids / Q000494:pharmacology; D017306:Annexin A2; D001980:Bronchi / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D002227:Carbazoles; D002467:Cell Nucleus / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000378:metabolism*; D005799:Genes, Dominant; D017930:Genes, Reporter; D016758:Genes, jun / Q000235:genetics; D015151:Immunoblotting; D007211:Indoles; D009154:Mutation; D009569:Nitric Oxide / Q000378:metabolism*; D009589:Nitrogen Oxides; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D009418:S100 Proteins; D013096:Spermine / Q000031:analogs & derivatives* / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014162:Transfection; D000099248:S100 Calcium Binding Protein A10
D000470:Alkaloids; D017306:Annexin A2; D002135:Calcium-Binding Proteins; D002227:Carbazoles; D015500:Chloramphenicol O-Acetyltransferase; D006152:Cyclic GMP; D017869:Cyclic GMP-Dependent Protein Kinases; D004791:Enzyme Inhibitors; D007211:Indoles; D009569:Nitric Oxide; D009589:Nitrogen Oxides; D012333:RNA, Messenger; D009418:S100 Proteins; D013096:Spermine; D013873:Thionucleotides; D018808:Transcription Factor AP-1; D000099248:S100 Calcium Binding Protein A10; C073601:KT 5823; C091861:spermine nitric oxide complex; C083763:8-bromoguanosino-3',5'-cyclic monophosphorothioate; C016276:8-bromocyclic GMP