TY - JOUR
T1 - Cloning and regulation of rat tissue inhibitor of metalloproteinases-2 in osteoblastic cells
AU - Cook, Thomas F.
AU - Burke, James S.
AU - Bergman, Kimberly D.
AU - Quinn, Cheryl O.
AU - Jeffrey, John J.
AU - Partridge, Nicola C.
PY - 1994/6
Y1 - 1994/6
N2 - 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.
AB - 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.
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U2 - 10.1006/abbi.1994.1243
DO - 10.1006/abbi.1994.1243
M3 - Article
C2 - 8203893
AN - SCOPUS:0028237787
SN - 0003-9861
VL - 311
SP - 313
EP - 320
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 2
ER -