Induction of Collagenase-3 (MMP-13) Expression in Human Skin Fibroblasts by Three-dimensional Collagen Is Mediated by p38 Mitogen-activated Protein Kinase

Abstract

Collagenase-3 (matrix metalloproteinase-13, MMP-13) is a recently identified human MMP with an exceptionally wide substrate specificity and restricted tissue-specific expression. Here we show that MMP-13 expression is induced in normal human skin fibroblasts cultured within three-dimensional collagen gel resulting in production and proteolytic activation of MMP-13. Induction of MMP-13 mRNAs by collagen gel was potently inhibited by blocking antibodies against ␣ 1 and ␣ 2 integrin subunits and augmented by activating antibody against ␤ 1 integrin subunit, indicating that both ␣ 1 ␤ 1 and ␣ 2 ␤ 1 integrins mediate the MMP-13-inducing cellular signal generated by three-dimensional collagen. Collagen-related induction of MMP-13 expression was dependent on tyrosine kinase activity, as it was abolished by treatment of fibroblasts with tyrosine kinase inhibitors genistein and herbimycin A. Contact of fibroblasts to three-dimensional collagen resulted in simultaneous activation of mitogen-activated protein kinases (MAPKs) in three distinct subgroups: extracellular signal-regulated kinase (ERK)1 and ERK2, Jun N-terminal kinase/ stress-activated protein kinase, and p38. Induction of MMP-13 expression was inhibited by treatment of fibroblasts with a specific p38 inhibitor, SB 203580, whereas blocking the ERK1,2 pathway (Raf/MEK1,2/ERK1,2) by PD 98059, a selective inhibitor of MEK1,2 activation potently augmented MMP-13 expression. Furthermore, specific activation of ERK1,2 pathway by 12-O-tetradecanoylphorbol-13-acetate markedly suppressed MMP-13 expression in dermal fibroblasts in collagen gel. These results show that collagen-dependent induction of MMP-13 in dermal fibroblasts requires p38 activity, and is inhibited by activation of ERK1,2. Therefore, the balance between the activity of ERK1,2 and p38 MAPK pathways appears to be crucial in regulation of MMP-13 expression in dermal fibroblasts, suggesting that p38 MAPK may serve as a target for selective inhibition of collagen degradation, e.g. in chronic dermal ulcers. Controlled degradation of extracellular matrix (ECM)1 is essential in physiologic situations involving connective tissue remodeling, such as tissue morphogenesis, angiogenesis, and tissue repair. On the other hand, excessive breakdown of connective tissue plays an important role in the pathogenesis of, e.g., rheumatoid arthritis, osteoarthritis, atherosclerosis, periodontitis, autoimmune blistering disorders of skin, and dermal photoaging, as well as in invasion and metastasis of tumor cells (see Refs. 1 and 2). Matrix metalloproteinases (MMPs) are a family of structurally related zinc-dependent endopeptidases collectively capable of degrading essentially all ECM components, and they apparently play an important role in ECM remodeling in the physiologic and pathologic situations mentioned above. At present, human MMP gene family contains 16 members, which can be divided into subgroups of collagenases, gelatinases, stromelysins, membrane-type MMPs, and novel MMPs based on their structure and substrate speci...

Clustering Induces a Lateral Redistribution of α2β1 Integrin from Membrane Rafts to Caveolae and Subsequent Protein Kinase C-dependent Internalization

“…For EV1 infections MOI 100 was used. have previously shown that mitogen activated protein kinases (MAPKs) are regulated by the collagen receptor integrins (Ravanti et al, 1999). Here, extracellular signal regulated kinase (ERK) was activated at very early time points after ␣2␤1 binding to EV1 ( Figure 5A).…”

“…Molecular Biology of the Cell 634 (Ivaska et al, 1999;Ravanti et al, 1999), the isoform of PKC (PKC ; Xu and Clark, 1997), and protein phosphatase 2A (PP2A; Ivaska et al, 2002a). Here, PKC␣ activity was needed for the internalization of ␣2␤1 integrin in caveolae.…”

p38 mitogen-activated kinase is a bidirectional regulator of human fibroblast collagenase-1 induction by three-dimensional collagen lattices

“…Recently, three-dimensional collagen lattices were demonstrated to stimulate MMP-13 and collagen gene expression via activation of p38 MAPK [18,19]. Thus we used SB203580, a specific inhibitor of p38 (p38α) and p38β [26], to study the role of p38 MAPK in the expression of MMP-1, another injury-related gene…”

“…Several studies have also reported that the p38 MAPK is required for the induction of MMP-1 gene expression by ceramide and tumour-derived extracellular matrix metalloproteinase inducer (EMMPRIN) [20,21]. Interestingly, although contracting collagen lattices induce both activation of p38 MAPK [18] and expression of MMP-1 [3,22] in human fibroblasts, the complete inhibition of p38 MAPK by SB203580 at 10 µM does not result in appreciable suppression of MMP-1 expression [18].…”

“…The p38 MAPK has been implicated in the signalling of type I collagen. For example, p38 MAPK activity is required for collagen-induced platelet aggregation [17] and for collagen-lattice-induced MMP-13 expression in fibroblasts [18]. Integrin α # β " , a collagen-binding integrin that regulates the contraction of collagen lattices, mediates p38-dependent collagen gene transcription [19].…”