The rabbit gene for 92-kDa matrix metalloproteinase. Role of AP1 and AP2 in cell type-specific transcription.
We isolated the rabbit gene for the 92-kDa matrix metalloproteinase, gelatinase B, and sequenced 1802 contiguous bases covering the first three exons and 522 bases of DNA upstream of the start site for transcription. The DNA between bases -519 and +19 is sufficient to drive expression of a reporter gene in early passage cultures of corneal fibroblasts or primary cultures of corneal epithelial cells. Basal activity of the gelatinase B promoter in fibroblasts is lower than a collagenase promotor of 1800 base pairs, but activity of both promotors is similarly stimulated by treatment of transfected cells with phorbol 12-myristate 13-acetate, and stimulation is enhanced by co-treatment with transforming growth factor-beta. In contrast, basal activity of the gelatinase B promotor in epithelial cells is higher than the collagenase promotor. Deletion analysis demonstrated that sequences upstream of base -330 confer cell type-specific activity to the gelatinase B promotor. Site-directed mutagenesis revealed that an AP1-like element within this region is specifically utilized by fibroblasts. This region also contains elements that confer the capacity for activation by AP2, a transcription factor found to be expressed by corneal epithelial cells but not by corneal fibroblasts. In contrast, AP2 does not activate the collagenase promotor. These results provide a molecular basis for the unique cell type-specific expression pattern of gelatinase B as compared to other matrix metalloproteinases.
MGH/Harvard Cutaneous Biology Research Center, Massachusetts General Hospital, Charlestown 02129.
M E
ME
Fini
J D
JD
Bartlett
M
M
Matsubara
W B
WB
Rinehart
M K
MK
Mody
M T
MT
Girard
M
M
Rainville
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D017364:Collagenases / Q000235:genetics*; D003315:Cornea / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D020780:Matrix Metalloproteinase 9; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011817:Rabbits; D016415:Sequence Alignment; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000502:physiology*; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic
D017931:DNA Primers; D004268:DNA-Binding Proteins; D018808:Transcription Factor AP-1; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D017364:Collagenases; D020780:Matrix Metalloproteinase 9; D013755:Tetradecanoylphorbol Acetate