Expression of the protein product of the prostaglandin synthase-2/TIS10 gene in mitogen-stimulated Swiss 3T3 cells

• Published in: The Journal of biological chemistry Vol. 268; no. 8; pp. 5425 - 5430
• Main Authors: KUJUBU, D. A, REDDY, S. T, FLETCHER, B. S, HERSCHMAN, H. R
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 15.03.1993
• Subjects: 3T3 Cells; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cell Line, Transformed; Chlorocebus aethiops; Cyclooxygenase Inhibitors - pharmacology; Dexamethasone - pharmacology; Enzyme Induction; Enzymes and enzyme inhibitors; expression; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; localization; Mice; Mitogens; Molecular Sequence Data; Oxidoreductases; prostaglandin synthase 2; Prostaglandin-Endoperoxide Synthases - biosynthesis; Prostaglandin-Endoperoxide Synthases - genetics; Prostaglandin-Endoperoxide Synthases - immunology; Swiss 3T3 cells; synthesis; Tetradecanoylphorbol Acetate - pharmacology; Vertebrata; Immunoprecipitation reaction; Mammalia; Mouse; Enzyme; Isozyme; Rodentia; 3T3-Cell line; Mitogen; Gene expression; Immunofluorescence; Prostaglandin synthase
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)53338-X

TIS10/PGS-2 encodes a prostaglandin synthase (PGS) distinct from the previously described enzyme PGS-1 (EC 1.14.99.1). We have now generated antipeptide antisera, directed to an amino acid sequence unique to the murine TIS10/PGS-2 protein, which specifically recognize the TIS10/PGS-2 antigen. TIS10/PGS-2 protein was undetectable in quiescent Swiss 3T3 cells. The level of TIS10/PGS-2 protein peaked between 6 and 8 h following phorbol ester stimulation of cells, then declined to basal levels after 18-24 h. Synthesis of TIS10/PGS-2 protein was dramatically increased in the second hour following mitogen stimulation and remained elevated for several hours. The half-life of the TIS10/PGS-2 protein was 4 h. Immunofluorescence studies demonstrated a perinuclear and cytoplasmic localization of the TIS10/PGS-2 antigen. As expected, detection of induced TIS10/PGS-2 antigen was dependent on protein synthesis. Metabolically labeled TIS10/PGS-2 protein migrated as a 71/73-kDa doublet following immunoprecipitation. Dexamethasone blocked both the TPA- and serum-induced appearance of TIS10/PGS-2 antigen. These studies demonstrate the existence of a mitogen-inducible, glucocorticoid-inhibitable, immunologically distinct prostaglandin synthase protein.