Nucleic Acids Research · 2001 · 262 citations · 30 references
Life SciencesCell BiologyCell SignalingSignal TransductionTranscriptional RegulationCartilage BreakdownCartilage MetabolismCytokine BiologyCytokine SignalingInflammatory SignalingConstitutive Expression SystemsConstitutive Gene ExpressionGene-specific Transcriptional RegulationNf-kappa B-dependent TranscriptionPromotersRunx2 Transcription FactorTissue-specific Transcription FactorsType Ii Collagen Degradation
Osteoarthritic chondrocytes secrete matrix metalloproteinase-13 (MMP-13) in response to interleukin-1 (IL-1), causing digestion of type II collagen in cartilage. Using chondrocytic cells, we previously determined that IL-1 induced a strong MMP-13 transcriptional response that requires p38 MAPK, JNK and the transcription factor NF-kappaB. Now, we have studied the tissue-specific transcriptional regulation of MMP-13. Constitutive expression of the transcription factor Runx-2 correlated with the ability of a cell type to express MMP-13 and was required for IL-1 induction; moreover, Runx-2 enhanced IL-1 induction of MMP-13 transcription by synergizing with the p38 MAPK signaling pathway. Transiently transfected MMP-13 promoters were not IL-1 inducible. However, -405 bp of stably integrated promoter was sufficient for 5- to 6-fold IL-1 induction of reporter activity and this integrated reporter required the same p38 MAPK pathway as the endogenous gene. Finally, mutation of the proximal Runx binding site and the proximal AP-1 site blunted the transcriptional response to IL-1, and double mutation synergistically decreased reporter activity. In summary, our data suggest that the transcriptional MMP-13 response to IL-1 is controlled by the p38 pathway interacting at the MMP-13 promoter through the tissue-specific transcription factor Runx-2 and the ubiquitous AP-1 transcription factor.
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Site-directed mutagenesis by overlap extension using the polymerase chain reaction
Steffan N. Ho, Henry D. Hunt, Robert M. Horton et al. · Gene · 1989 · 7.5K citations
Osf2/Cbfa1: A Transcriptional Activator of Osteoblast Differentiation
Patricia Ducy, Rui Zhang, Valérie Geoffroy et al. · Cell · 1997 · 4.3K citations · Full text