Alternate COX-2 transcripts are differentially regulated: implications for post-transcriptional control

Biochem Biophys Res Commun. 1997 May 8;234(1):85-9. doi: 10.1006/bbrc.1997.6586.

Abstract

Prostaglandin (PG) synthesis during inflammation occurs mainly via the transcriptionally regulated cyclooxygenase, COX-2. In pulmonary type II A549 cells, Northern analysis identified multiple IL-1 beta-inducible COX-2 mRNA transcripts. Amplification of 3'-cDNA ends by anchored PCR revealed products corresponding to the predominant 4.5 and 2.7 kb transcripts. Sequence analysis of amplification products indicated that these transcripts arose by alternate consensus and non-consensus polyadenylation site usage. The predominant 4.5 kb transcript showed a half-life in excess of two hours that was further stabilized by IL-1 beta. In addition, the COX-2 3'-untranslated region (UTR), which contains 22 copies of the putative RNA instability motif, AUUUA, when cloned downstream of a constitutively expressed luciferase gene, was found to confer partial IL-1 beta responsiveness in LA-4 cells. Finally, in vivo in LPS-treated rats, differential expression of similar COX-2 mRNA isoforms was also observed. Taken together these data suggest a functional role for post-transcriptional mechanisms, including alternate polyadenylation, in the control of COX-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Northern
  • Cells, Cultured
  • Cyclooxygenase 2
  • Genes, Reporter
  • Humans
  • Interleukin-1 / pharmacology
  • Isoenzymes / genetics*
  • Lipopolysaccharides / pharmacology
  • Luciferases / genetics
  • Male
  • Membrane Proteins
  • Mice
  • Poly A / genetics
  • Polymerase Chain Reaction
  • Prostaglandin-Endoperoxide Synthases / genetics*
  • RNA Processing, Post-Transcriptional / genetics*
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism*
  • Rats
  • Rats, Wistar
  • Transcription, Genetic

Substances

  • Interleukin-1
  • Isoenzymes
  • Lipopolysaccharides
  • Membrane Proteins
  • RNA, Messenger
  • Poly A
  • Luciferases
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases