Tissue inhibitor of metalloproteinases-3 (TIMP-3) is an extracellular matrix-associated protein with a distinctive pattern of expression in mouse cells and tissues

• Published in: The Journal of biological chemistry Vol. 269; no. 12; pp. 9352 - 9360
• Main Authors: LECO, K. J, KHOKHA, R, PAVLOFF, N, HAWKES, S. P, EDWARDS, D. R
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 25.03.1994
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Base Sequence; Biological and medical sciences; Cell Nucleus - metabolism; Cloning, Molecular; Dexamethasone - pharmacology; Enzymes and enzyme inhibitors; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; Genes; Hydrolases; Mice; Molecular Sequence Data; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Restriction Mapping; RNA, Messenger - genetics; Sequence Alignment; Sequence Homology, Amino Acid; Tetradecanoylphorbol Acetate - pharmacology; Tissue Distribution; Tissue Inhibitor of Metalloproteinase-3; Transcription, Genetic; Transforming Growth Factor beta - pharmacology; Enzyme; Rodentia; Enzyme inhibitor; Tissue specificity; Metalloendopeptidases; Primary structure; Gene expression; Vertebrata; Mammalia; Mouse; Hydrolases; Extracellular matrix; Molecular cloning; Proteinases
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)37115-6

We have isolated cDNA clones corresponding to a novel mouse metalloproteinase inhibitor. Five overlapping cDNA clones contain most of the information for a prominent 4.5-kilobase transcript that was detected in RNA from mouse fibroblasts and adult tissues. Sequence analysis revealed an open reading frame (ORF) for a protein of 212 amino acids that is 80% identical to chicken inhibitor of metalloproteinases-3 (ChIMP-3). The 3'-untranslated sequence also showed remarkable conservation with the chicken gene. The ORF directed the expression of a 24-kDa protein in COS-1 cells that localized to the extracellular matrix (ECM). On the basis of these similarities we propose to identify the new gene as murine tissue inhibitor of metalloproteinases-3 (TIMP-3). Mouse C3H 10T1/2 fibroblasts produced a 24-kDa metalloproteinase inhibitor that also localized to the ECM and was recognized by a polyclonal antibody to ChIMP-3. Like TIMP-1, TIMP-3 was highly inducible in mouse C3H 10T1/2 fibroblasts by phorbol ester (PMA), epidermal growth factor (EGF), and transforming growth factor-beta 1, but nuclear run-on assays showed that the on/off transcription kinetics were faster for TIMP-3 than TIMP-1. A major difference in vitro was the stimulation of expression of TIMP-3 by dexamethasone which inhibits EGF- and PMA-induced TIMP-1 transcription. Also, TIMP-3 showed a distinctive pattern of expression in adult tissues with abundant transcripts detected in kidney, lung, and brain but only low levels detected in bone, a prominent location of TIMP-1 transcripts. We propose that TIMP-3 functions in a tissue-specific fashion as part of an acute response to remodeling stimuli.