A composite element binding the vitamin D receptor and the retinoic X receptor alpha mediates the transforming growth factor-beta inhibition of decorin gene expression in articular chondrocytes.
Decorin, a small leucine-rich proteoglycan may play an important role in the attempt of cartilage repair initiated by chondrocytes in early stages of osteoarthritis, through its ability to bind collagen fibrils and growth factors such as transforming growth factor-beta (TGF-beta). We previously demonstrated that TGF-beta decreased decorin mRNA steady state levels in articular chondrocytes (Demoor, M., Rédini, F., Boittin, M., and Pujol, J.-P. (1998) Biochim. Biophys. Acta 1398, 179-191). Here, we investigated the effect of TGF-beta on decorin gene expression in both primary cultures of articular chondrocytes and chondrocytes dedifferentiated by serial passages. Transient transfection of cells with plasmid constructs of the decorin promoter linked to the luciferase reporter gene revealed transcriptional repression by TGF-beta, in fully differentiated as well as dedifferentiated chondrocytes. Experiments with 5'-deleted constructs allowed characterization of a TGF-beta-responsive element in the shortest construct (base pairs (bp) -155/+269). DNase I footprinting analysis delineated a negative TGF-beta-responsive region between -140 and -111 bp in the decorin proximal promoter. Gel retardation assays demonstrated that TGF-beta modulates decorin gene expression through transcription factors, the nature and mode of action of which depend on the differentiation state of the chondrocytes; two DNA-protein complexes were formed in the region -144/-127 bp with nuclear extracts from primary chondrocytes, whereas a higher mobility complex was observed in the -127/-111 bp region for dedifferentiated cells. Antibodies against vitamin D and retinoic acid receptors used in supershift experiments showed that these nuclear receptors are involved in the regulation of decorin gene expression in articular chondrocytes.
Demoor-Fossard
M
M
Laboratoire de Biochimie du Tissu Conjonctif, Faculté de Médecine, niv. 3, 14032 Caen cedex, France.
Galéra
P
P
Santra
M
M
Iozzo
R V
RV
Pujol
J P
JP
Rédini
F
F
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000502:physiology; D018983:DNA Footprinting; D058575:Decorin; D003850:Deoxyribonuclease I / Q000378:metabolism; D004586:Electrophoresis; D016326:Extracellular Matrix Proteins; D005786:Gene Expression Regulation / Q000187:drug effects*; D020868:Gene Silencing; D006801:Humans; D008233:Lymphotoxin-alpha / Q000378:metabolism / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011509:Proteoglycans / Q000096:biosynthesis / Q000235:genetics*; D011817:Rabbits; D018167:Receptors, Calcitriol / Q000502:physiology*; D018168:Receptors, Retinoic Acid / Q000502:physiology*; D047488:Retinoid X Receptors; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic
C546878:DCN protein, human; D058575:Decorin; D016326:Extracellular Matrix Proteins; D008233:Lymphotoxin-alpha; D011509:Proteoglycans; D018167:Receptors, Calcitriol; D018168:Receptors, Retinoic Acid; D047488:Retinoid X Receptors; D014157:Transcription Factors; D003850:Deoxyribonuclease I