Induction of secreted type IIA phospholipase A2 gene transcription by interleukin-1beta. Role of C/EBP factors

J Biol Chem. 2000 Jul 28;275(30):22686-94. doi: 10.1074/jbc.M001250200.

Abstract

Secreted type IIA phospholipase A(2), which is involved in arachidonic acid release, is abundantly produced by chondrocytes and secreted in the synovial fluids of patients affected by rheumatoid arthritis. Transfection experiments showed that interleukin-1beta stimulates the phospholipase A(2) [-1614; +20] promoter activity by 6-7-fold and that the [-210; -176] fragment is critical for this stimulation. CAAT enhancer-binding protein (C/EBP) beta and C/EBPdelta transcription factors bind to this element as shown by bandshift experiments. Interleukin-1beta increased the levels of C/EBPdelta mRNA as soon as 2 h and up to 24 h without affecting those of C/EBPbeta. Higher amounts of C/EBPdelta proteins correlate with the stimulation of C/EBPdelta mRNA. Mutations or 5' deletions in the upstream [-247; -210] region reduced by 2-fold the basal and interleukin-1beta-stimulated transcription activities. Two types of factors bind to overlapping sequences on this fragment: NF1-like proteins and the glucocorticoid receptor. The glucocorticoid receptor is responsible for a moderate stimulation of the promoter activity by dexamethasone and may interact with C/EBP factors to achieve a full transcription activity in basal conditions and in the presence of interleukin-1beta. A [-114; -85] proximal regulatory element forms three complexes in bandshift experiments, the slowest mobility one involving the Sp1 zinc finger factor. Mutation of this sequence reduced to 2-fold the stimulation of the promoter activity by interleukin-1beta or the C/EBP factors. Induction of the transcription of secreted type IIA phospholipase A(2) gene by interleukin-1beta in chondrocytes absolutely requires C/EBPbeta and C/EBPdelta factors but does not involve NF-kappaB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • CCAAT-Enhancer-Binding Proteins
  • Cells, Cultured
  • Chondrocytes / cytology
  • Chondrocytes / enzymology
  • Chondrocytes / metabolism
  • DNA Primers
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology
  • Female
  • Gene Expression Regulation, Enzymologic / physiology*
  • Interleukin-1 / physiology*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Nuclear Proteins / physiology
  • Phospholipases A / genetics*
  • Phospholipases A2
  • Promoter Regions, Genetic
  • Protein Binding
  • Rabbits
  • Receptors, Glucocorticoid / metabolism
  • Regulatory Sequences, Nucleic Acid
  • Sp1 Transcription Factor / metabolism
  • Transcription, Genetic / physiology*

Substances

  • CCAAT-Enhancer-Binding Proteins
  • DNA Primers
  • DNA-Binding Proteins
  • Interleukin-1
  • Nuclear Proteins
  • Receptors, Glucocorticoid
  • Sp1 Transcription Factor
  • Phospholipases A
  • Phospholipases A2