Interferon-gamma- and phorbol myristate acetate-responsive elements involved in intercellular adhesion molecule-1 mRNA stabilization.
Treatment of cells with interferon (IFN)-gamma or phorbol myristate acetate (PMA) induces up-regulation of the level of intercellular adhesion molecule-1 (ICAM-1; CD54) mRNA by stabilization of an otherwise labile mRNA. Here, we have generated various deletion mutants of ICAM-1 and stably transfected them into the murine fibroblast Ltk- cells that express no endogenous ICAM-1 or -2 (CD102) in an effort to define the regions within ICAM-1 mRNA responsive to IFN-gamma or PMA. Induction of ICAM-1 mRNA in the transfected L cells by the treatment with IFN-gamma revealed that the truncation of the region of ICAM-1 mRNA encoding the cytoplasmic domain made it non-responsive to IFN-gamma whereas all other regions were dispensable. In contrast, PMA-induced accumulation of ICAM-1 mRNA required the 3'-untranslated region (UTR). To further elucidate the role of these regions in mRNA destabilization and responsiveness to IFN-gamma and PMA, ICAM-2 mRNA that is stable and not responsive to IFN-gamma or PMA was used as a reporter gene. The putative IFN-gamma-responsive region of ICAM-1 mRNA encoding its cytoplasmic domain rendered it unstable and responsive to IFN-gamma but not PMA. Conversely, the 3'-UTR of ICAM-1 fused with ICAM-2 mRNA also made it unstable and responsive to PMA but not IFN-gamma. Half-life analysis showed that the induction of these chimeric mRNAs by IFN-gamma and PMA was due, at least in part, to the prolongation of their turnover rate. These results taken together demonstrate that two distinct regions of ICAM-1 mRNA regulate its stability, one encoding the cytoplasmic domain and responsive to IFN-gamma and the other in the 3'-UTR and responsive to PMA.
Ohh
M
M
Department of Medical Genetics, University of British Columbia, Vancouver, Canada.
Takei
F
F
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D005822:Genetic Vectors; D006207:Half-Life; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis* / Q000235:genetics; D007371:Interferon-gamma / Q000494:pharmacology*; D007700:Kinetics; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism*; D017384:Sequence Deletion; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D018799:Intercellular Adhesion Molecule-1; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate