A novel role for GADD45beta as a mediator of MMP-13 gene expression during chondrocyte terminal differentiation.
The growth arrest and DNA damage-inducible 45beta (GADD45beta) gene product has been implicated in the stress response, cell cycle arrest, and apoptosis. Here we demonstrated the unexpected expression of GADD45beta in the embryonic growth plate and uncovered its novel role as an essential mediator of matrix metalloproteinase-13 (MMP-13) expression during terminal chondrocyte differentiation. We identified GADD45beta as a prominent early response gene induced by bone morphogenetic protein-2 (BMP-2) through a Smad1/Runx2-dependent pathway. Because this pathway is involved in skeletal development, we examined mouse embryonic growth plates, and we observed expression of Gadd45beta mRNA coincident with Runx2 protein in pre-hypertrophic chondrocytes, whereas GADD45beta protein was localized prominently in the nucleus in late stage hypertrophic chondrocytes where Mmp-13 mRNA was expressed. In Gadd45beta(-/-) mouse embryos, defective mineralization and decreased bone growth accompanied deficient Mmp-13 and Col10a1 gene expression in the hypertrophic zone. Transduction of small interfering RNA-GADD45beta in epiphyseal chondrocytes in vitro blocked terminal differentiation and the associated expression of Mmp-13 and Col10a1 mRNA in vitro. Finally, GADD45beta stimulated MMP-13 promoter activity in chondrocytes through the JNK-mediated phosphorylation of JunD, partnered with Fra2, in synergy with Runx2. These observations indicated that GADD45beta plays an essential role during chondrocyte terminal differentiation.
Ijiri
Kosei
K
Beth Israel Deaconess Medical Center, New England Baptist Bone and Joint Institute, and Harvard Medical School, Boston, Massachusetts 02115, USA.
Zerbini
Luiz F
LF
Peng
Haibing
H
Correa
Ricardo G
RG
Lu
Binfeng
B
Walsh
Nicole
N
Zhao
Yani
Y
Taniguchi
Noboru
N
Huang
Xu-Ling
XL
Otu
Hasan
H
Wang
Hong
H
Wang
Jian Fei
JF
Komiya
Setsuro
S
Ducy
Patricia
P
Rahman
Mahboob U
MU
Flavell
Richard A
RA
Gravallese
Ellen M
EM
Oettgen
Peter
P
Libermann
Towia A
TA
Goldring
Mary B
MB
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D001846:Bone Development; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D001842:Bone and Bones / Q000378:metabolism; D002356:Cartilage / Q000378:metabolism; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D017364:Collagenases / Q000096:biosynthesis*; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D004249:DNA Damage; D005269:Femur / Q000378:metabolism; D051921:Fos-Related Antigen-2 / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D017403:In Situ Hybridization; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D008156:Luciferases / Q000378:metabolism; D053509:Matrix Metalloproteinase 13; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D051898:Smad1 Protein / Q000378:metabolism; D013977:Tibia / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism; D015854:Up-Regulation; D000097642:GADD45 Proteins
D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D017364:Collagenases; D050659:Core Binding Factor Alpha 1 Subunit; D051921:Fos-Related Antigen-2; D047908:Intracellular Signaling Peptides and Proteins; D008156:Luciferases; D053509:Matrix Metalloproteinase 13; D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051898:Smad1 Protein; D016212:Transforming Growth Factor beta; D000097642:GADD45 Proteins; C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; C497872:FOSL2 protein, human; C507079:MMP13 protein, human; C507081:Mmp13 protein, mouse