Cell type-specific regulation of SL-1 and SL-2 genes. Induction of the SL-2 gene but not the SL-1 gene by human keratinocytes in response to cytokines and phorbolesters.
The stromelysin-2 (SL-2) gene is transcriptionally active in normal human keratinocytes and encodes a secreted, catalytically competent but latent matrix metalloproteinase. Phorbolester induction resulted in the emergence of SL-2 (but not SL-1 transcripts), whereas the opposite was true for human mucosal fibroblasts. Expression of keratinocyte SL-2 was also induced by the two keratinocyte growth factors, transforming growth factor-alpha and epidermal growth factor, by the proinflammatory cytokine, tumor necrosis factor-alpha, but, somewhat surprisingly, not by interleukin-1 beta. The latent SL-2 proenzyme was isolated from 12-O-tetradecanoylphorbol-13-acetate-induced keratinocytes by immunoaffinity chromatography using a cross-reactive antibody raised against human SL-1. This procedure led to the recovery of a single M(r) 54,000 molecular species at a level of approximately 0.2 microgram/ml of culture medium. Amino-terminal sequencing identified the protein as SL-2 and verified the predicted signal sequence cleavage site. Conformational activation of latent SL-2 precursor by SDS gave rise to a full-length, uncleaved (M(r) 54,000) active form and at the same time exposed a cryptic thiol group. By contrast, organomercurial activation resulted in autolytic truncation of the molecule with loss of M(r) approximately 10,000 propeptide. SL-2 shared with (human fibroblast) SL-1 the ability to cleave casein, to "superactivate" fibroblast type procollagenase, and to form apparently binary, SDS-resistant complexes with tissue inhibitor of metalloproteinases-1.
Windsor
L J
LJ
Department of Oral Biology, University of Alabama School of Dentistry, Birmingham.
Grenett
H
H
Birkedal-Hansen
B
B
Bodden
M K
MK
Engler
J A
JA
Birkedal-Hansen
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D004247:DNA; D004790:Enzyme Induction / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000201:enzymology*; D053506:Matrix Metalloproteinase 10; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D016207:Cytokines; D006023:Glycoproteins; D004247:DNA; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D053506:Matrix Metalloproteinase 10; D013755:Tetradecanoylphorbol Acetate