p11 expression in human bronchial epithelial cells is increased by nitric oxide in a cGMP-dependent pathway involving protein kinase G activation

J Biol Chem. 2001 Nov 30;276(48):44613-21. doi: 10.1074/jbc.M104993200. Epub 2001 Sep 24.

Abstract

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.

MeSH terms

  • Alkaloids / pharmacology
  • Annexin A2*
  • Bronchi / metabolism*
  • Calcium-Binding Proteins / biosynthesis*
  • Calcium-Binding Proteins / metabolism*
  • Carbazoles*
  • Cell Nucleus / metabolism
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Cyclic GMP / analogs & derivatives*
  • Cyclic GMP / metabolism*
  • Cyclic GMP / pharmacology
  • Cyclic GMP-Dependent Protein Kinases / antagonists & inhibitors
  • Cyclic GMP-Dependent Protein Kinases / metabolism*
  • Dose-Response Relationship, Drug
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Epithelial Cells / metabolism*
  • Genes, Dominant
  • Genes, Reporter
  • Genes, jun / genetics
  • Immunoblotting
  • Indoles*
  • Mutation
  • Nitric Oxide / metabolism*
  • Nitrogen Oxides
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA, Messenger / metabolism
  • S100 Proteins*
  • Spermine / analogs & derivatives*
  • Spermine / pharmacology
  • Thionucleotides / pharmacology
  • Time Factors
  • Transcription Factor AP-1 / metabolism
  • Transfection

Substances

  • Alkaloids
  • Annexin A2
  • Calcium-Binding Proteins
  • Carbazoles
  • Enzyme Inhibitors
  • Indoles
  • Nitrogen Oxides
  • RNA, Messenger
  • S100 Proteins
  • S100 calcium binding protein A10
  • Thionucleotides
  • Transcription Factor AP-1
  • KT 5823
  • spermine nitric oxide complex
  • 8-bromoguanosino-3',5'-cyclic monophosphorothioate
  • Spermine
  • 8-bromocyclic GMP
  • Nitric Oxide
  • Chloramphenicol O-Acetyltransferase
  • Cyclic GMP-Dependent Protein Kinases
  • Cyclic GMP