Cloning and Regulation of Rat Tissue Inhibitor of Metalloproteinases-2 in Osteoblastic Cells

T. F. Cook, J. S. Burke, K. D. Bergman, C. O. Quinn, J. J. Jeffrey, Nicola Partridge

Research output: Contribution to journalArticle

Abstract

Rat tissue inhibitor of metalloproteinases-2 (TIMP-2) was cloned from a UMR 106-01 rat osteoblastic osteosarcoma cDNA library. The 969-bp full-length clone demonstrates 98 and 86% sequence identity to human TIMP-2 at the amino acid and nucleic acid levels, respectively. Parathyroid hormone (PTH), at 10-8 M, stimulates an approximately twofold increase in both the 4.2-and 1.0-kb transcripts over basal levels in UMR cells after 24 h of exposure. The PTH stimulation of TIMP-2 transcripts was not affected by the inhibitor of protein synthesis, cycloheximide (10-5 M), suggesting a primary effect of the hormone. This is in contradistinction to regulation of interstitial collagenase (matrix metalloproteinase-l) by PTH in these same cells. Nuclear run-on assays demonstrate that PTH causes an increase in TIMP-2 transcription that parallels the increase in message levels. Parathyroid hormone, in its stimulation of TIMP-2 mRNA, appears to act through a signal transduction pathway involving protein kinase A (PRA) since the increase in TIMP-2 mRNA is reproduced by treatment with the cAMP analogue, 8-bromo-cAMP (5 × 10-3 M). The protein kinase C and calcium pathways do not appear to be involved due to the lack of effect of phorbol l2-myristate 13-acetate (2.6 × 10-6 M) and the calcium ionophore, ionomycin (10-7 M), on TIMP-2 transcript abundance. In this respect, regulation of TIMP-2 and collagenase in osteoblastic cells by PTH are similar. However, we conclude that since stimulation of TIMP-2 transcription is a primary event, the PRA pathway must be responsible for a direct increase in transcription of this gene.

Original languageEnglish (US)
Pages (from-to)313-320
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume311
Issue number2
DOIs
StatePublished - Jun 1994

Fingerprint

Tissue Inhibitor of Metalloproteinase-2
Cloning
Rats
Organism Cloning
Parathyroid Hormone
Transcription
Cyclic AMP-Dependent Protein Kinases
8-Bromo Cyclic Adenosine Monophosphate
Signal transduction
Messenger RNA
Matrix Metalloproteinase 1
Ionomycin
Protein Synthesis Inhibitors
Calcium Ionophores
Myristic Acid
Collagenases
Osteosarcoma
Cycloheximide
Matrix Metalloproteinases
Gene Library

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

Cloning and Regulation of Rat Tissue Inhibitor of Metalloproteinases-2 in Osteoblastic Cells. / Cook, T. F.; Burke, J. S.; Bergman, K. D.; Quinn, C. O.; Jeffrey, J. J.; Partridge, Nicola.

In: Archives of Biochemistry and Biophysics, Vol. 311, No. 2, 06.1994, p. 313-320.

Research output: Contribution to journalArticle

Cook, T. F. ; Burke, J. S. ; Bergman, K. D. ; Quinn, C. O. ; Jeffrey, J. J. ; Partridge, Nicola. / Cloning and Regulation of Rat Tissue Inhibitor of Metalloproteinases-2 in Osteoblastic Cells. In: Archives of Biochemistry and Biophysics. 1994 ; Vol. 311, No. 2. pp. 313-320.
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