TY - JOUR
T1 - ADAMTS9 activation by interleukin 1β via NFATc1 in OUMS-27 chondrosarcoma cells and in human chondrocytes
AU - Yaykasli, Kursat Oguz
AU - Oohashi, Toshitaka
AU - Hirohata, Satoshi
AU - Hatipoglu, Omer Faruk
AU - Inagawa, Kiichi
AU - Demircan, Kadir
AU - Ninomiya, Yoshifumi
N1 - Funding Information:
Fig. 6 Effect of IL-1β on ADAMTS9 promoter activity in OUMS-27 and in chondrocytes. a A schematic representation of sequential 5′ deletion of ADAMTS9 promoter. 5′ deletion fragments of ADAMTS9 promoter were generated by PCR and were ligated in the upstream of the Secreted Metridia longa luciferase gene in the pMetLuc-Reporter vector. The location of the NBEs is shown. b Relative Luciferase Activity. OUMS-27 and chondrocytes were transfected with three promoter deletion constructs. After 24 h, the medium was collected, and then replaced to the fresh medium containing IL-1β. Secreted Luciferase activity in medium was measured 24 h post-cytokine treatment. The relative secreted luciferase activity in the medium was calculated by the ratio of cytokines treatment medium to nontreatment medium. Luciferase activity were normalized to the activity of control (value = 1) and represent mean ± SD. Significance compared to the control *P \ 0.05 Acknowledgments This work was supported in part by a Grant-in-Aid for Scientific Research to T.O. (20659232) and S.H. (20390399) from the Japan Society for the Promotion of Science (JSPS) and a Research for Collaborative Development of Innovative Seeds from Japan Science and Technology Corporation (JST) to T.O. We thank Drs. Matsushita, Kondo and Sumiyoshi for technical advice and our colleagues at Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences for stimulating discussions.
PY - 2009
Y1 - 2009
N2 - ADAMTS9 is a member of the disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) genes, with aggrecan-degrading activity. It has also been characterized to be reactive and highly activated ADAMTS by IL-1β in both chondrosarcoma cells and human chondrocytes (Demircan et al. Arthritis Rheum 52:1451-1460, 2005). In order to understand the regulation of ADAMTS9 gene expression a functional 3.0 kb human ADAMTS9 promoter has been cloned and characterized. A sequence analysis of the promoter revealed the presence of putative binding sites for Nuclear Factor of Activated T cells (NFAT), which is commonly found in the ADAMTS4 and ADAMTS5 promoters. NFATc1 was up-regulated in an activated form by IL-1β in human chondrocytes. The IL-1β inducible ADAMTS9 expression was inhibited by NFAT inhibitors, FK506 and 11Arg (11R)-VIVIT. Furthermore, direct binding of NFATc1 on distal and proximal promoters of ADAMTS9 was demonstrated by a chromatin immunoprecipitation assay. Promoter-reporter assays supported those results. These findings may provide a better understanding of the regulation of ADAMTS9 expression induced by inflammatory cytokines.
AB - ADAMTS9 is a member of the disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) genes, with aggrecan-degrading activity. It has also been characterized to be reactive and highly activated ADAMTS by IL-1β in both chondrosarcoma cells and human chondrocytes (Demircan et al. Arthritis Rheum 52:1451-1460, 2005). In order to understand the regulation of ADAMTS9 gene expression a functional 3.0 kb human ADAMTS9 promoter has been cloned and characterized. A sequence analysis of the promoter revealed the presence of putative binding sites for Nuclear Factor of Activated T cells (NFAT), which is commonly found in the ADAMTS4 and ADAMTS5 promoters. NFATc1 was up-regulated in an activated form by IL-1β in human chondrocytes. The IL-1β inducible ADAMTS9 expression was inhibited by NFAT inhibitors, FK506 and 11Arg (11R)-VIVIT. Furthermore, direct binding of NFATc1 on distal and proximal promoters of ADAMTS9 was demonstrated by a chromatin immunoprecipitation assay. Promoter-reporter assays supported those results. These findings may provide a better understanding of the regulation of ADAMTS9 expression induced by inflammatory cytokines.
KW - ADAMTS
KW - Arthritis
KW - Chondrocyte
KW - NFAT
KW - Protein transduction domain
KW - VIVIT
UR - http://www.scopus.com/inward/record.url?scp=59949095681&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=59949095681&partnerID=8YFLogxK
U2 - 10.1007/s11010-008-9965-4
DO - 10.1007/s11010-008-9965-4
M3 - Article
C2 - 19052845
AN - SCOPUS:59949095681
SN - 0300-8177
VL - 323
SP - 69
EP - 79
JO - Enzymologia
JF - Enzymologia
IS - 1-2
ER -