Abstract
Mutations in human SOX9 are associated with campomelic dysplasia (CD), characterised by skeletal malformation and XY sex reversal1–3. During chondrogenesis in the mouse, Sox9 is co-expressed with Col2a1, the gene encoding type-ll collagen, the major cartilage matrix protein4. Col2a1 is therefore a candidate regulatory target of SOX9. Regulatory sequences required for chondrocyte-specific expression of the type-ll collagen gene have been localized to conserved sequences in the first intron in rats, mice and humans5–8. We show here that SOX9 protein binds specifically to sequences in the first intron of human COL2A1. Mutation of these sequences abolishes SOX9 binding and chondrocyte-specific expression of a COL2A1-driven reporter gene (COL2A1-lacZ) in transgenic mice. Furthermore, ectopic expression of Sox9 trans-activates both a COLZA1-driven reporter gene and the endogenous Col2a1 gene in transgenic mice. These results demonstrate that COL2A1 expression is directly regulated by SOX9 protein in vivo and implicate abnormal regulation of COL2A1 during chondrogenesis as a cause of the skeletal abnormalities associated with campomelic dysplasia.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Foster, J.W. et al. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 372, 525–530 (1994).
Wagner, T. et al. Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY-related gene SOX9. Cell 79, 1111–1120 (1994).
Schafer, A.J. et al. Campomelic dysplasia with XY sex reversal: diverse phenotypes resulting from mutations in a single gene. Ann. NYAcad. Sci. 785, 137–149 (1996).
Ng, L.J. . et al. SOX9 binds DNA, activates transcription and co-expresses with type II collagen during chondrogenesis in the mouse. Devl. Biol. 183, 108–121 (1997).
Wang, L, Balakir, R. & Horton, W.E. Identification of a cis-acting sequence in the collagen II enhancer required for chondrocyte expression and the binding of a chondrocyte nuclear factor. J. Biol. Chem. 266, 19878–19881 (1991).
Zhou, G. et al. A 182 bp fragment of the mouse proa1(ll) collagen gene is sufficient to direct chondrocyte expression in transgenic mice. J. Cell Sci. 108, 3677–3684 (1995).
Lefebvre, V. et al. A 18-base-pair sequence in the mouse proa 1(11) collagen gene is sufficient for expression in cartilage and binds nuclear proteins that are selectively expressed in chondrocytes. Mol. Cell. Biol. 16, 4512–4523 (1996).
Seghatoleslami, M.R. et al. Differential regulation of COL2A1 expression in developing and mature chondrocytes. Matrix Biol. 14, 753–764 (1994).
Harley, V.R., Lovell-Badge, R. & Goodfellow, P.N. Definition of a consensus DNA binding site for SRY. Nucleic Acids Res. 22, 1500–1501 (1994).
Denny, P., Swift, S., Connor, F. & Ashworth, A. SRY-related gene expressed during spermatogenesis in the mouse encodes a sequence-specific DNA binding protein. EMBO J. 11, 3705–3712 (1992).
van de Wetering, M., Oosterwegel, M., van Norren, K. & Clevers, H. Sox-4, an Sry-like HMG box protein, is a transcriptional activator in lymphocytes. EMBO J. 12, 3847–3854 (1993).
Hosking, B.M., Muscat, G.E.O., Koopman, P., Dowhan, D.H. & Dunn, T.L. Trans-activation and DNA-binding properties of the transcription factor, Sox-18. Nucl. Acids Res. 23, 2626–2628 (1995).
Kanai, Y. et al. Identification of two Sox17 messenger RNA isoforms with and without the high mobility group box region, and their differential expression in mouse spermatogenesis. J. Cell Biol. 133, 1–15 (1996).
Kent, J., Wheatley, S.C., Andrews, J.E., Sinclair, A.H. & Koopman, P. A male-specific role for SOX9 in vertebrate sex determination. Development 122, 2813–2822 (1996).
Sham, M.H. et al. The zinc finger gene Krox20 regulates HoxB2 (Hox2.8) during hindbrain segmentation. Cell 72, 183–196 (1993).
Kamachi, Y.S. et al. Involvement of SOX proteins in lens-specific activation of crystallin genes. EMBO J. 14, 1310–1319 (1995).
Yuan, H., Corbi, N., Basilico, C. & L Developmental-specific activity of the FGF-4 enhancer requires synergistic action of Sox2 and Oct-3. Genes Dev. 9, 2635–3645 (1995).
Chavrier, P. et al. The segment-specific gene Krox-20 encodes a transcription factor with binding sites in the promoter region of the Hox-1.4 gene. EMBO J. 9, 1209–1218 (1994).
Nonchev, S. et al. Segmental expression of Hoxa-2 in the hindbrain is directly regulated by Krox-20. Development 122, 543–554 (1995).
Carlsson, P., Waterman, M.L. & Jones, K.A. The hLEF-1/TCF-1a HMG protein contains a context-dependent transcriptional activation domain that induces the TCRa enhancer in T cells. Genes Dev. 7, 2418–2430 (1993).
Giese, K. & Grosschedl, R. LEF-1 contains an activation domain that stimulates transcription only in the context of factor-binding sites. EMBO J. 12, 4667–4676 (1993).
Pontiggia, A. et al. Sex-reversing mutations affect the architecture of SRY-DNA complexes. EMBO J. 13, 6115–6124 (1994).
Grosschedl, R., Giese, K. & Pagel, J. HMG domain proteins architectural elements in the assembly of nucleoprotein structures. Trends Genet. 10, 94–99 (1994).
Savagner, P. et al. Collagen II promoter and enhancer interact synergistically through Sp1 and distinct nuclear factors. DNA Cell Biol. 14, 501–510 (1995).
Li, S-W. et al. Transgenic mice with a targeted inactivation of the Col2a1 gene for collagen II develop a skeleton with membranous and periostea! bone but no endochondral bone. Genes Dev. 9, 2821–2830 (1995).
Chan, D., Cole, W.G., Chow, C.W., Mundlos, S. & Bateman, J.F. A COL2A1 mutation in achondrogenesis type II results in the replacement of type II collagen by type I and III collagens in cartilage. J. Biol. Chem. 270, 1747–1753 (1995).
Ritvaniemi, P. et al. Analysis of the COL2A1 gene detects mutations in over 20 percent of patients with chondrodysplasias and up to 2 percent of patients with familial osteoarthritis. Arthritis Rheum. 38, 999–1004 (1995).
Yee, S.-P. & Rigby, P.W.J. The regulation of myogenin gene expression during the embryonic development of the mouse. Genes Dev. 7, 1277–1289 (1993).
Ng, L.J., Tam, P.P.L. . & Cheah, K.S.E. Preferential expression of alternatively spliced mRNAs encoding type II procollagen with a cysteine-rich amino-propeptide in differentiating cartilage and nonchondrogenic tissues during early mouse development. Devl. Biol. 159, 403–417 (1993).
Wright, E.M. et al. The Sry-related gene Sox9 is expressed during chondrogenesis in mouse embryos. Nature Genet. 9, 15–20 (1995).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bell, D., Leung, K., Wheatley, S. et al. SOX9 directly regulates the type-ll collagen gene. Nat Genet 16, 174–178 (1997). https://doi.org/10.1038/ng0697-174
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/ng0697-174
This article is cited by
-
Inhibition of HOXD11 promotes cartilage degradation and induces osteoarthritis development
Journal of Orthopaedic Surgery and Research (2024)
-
Multi-omics analysis of sarcospan overexpression in mdx skeletal muscle reveals compensatory remodeling of cytoskeleton-matrix interactions that promote mechanotransduction pathways
Skeletal Muscle (2023)
-
Precise modulation of transcription factor levels identifies features underlying dosage sensitivity
Nature Genetics (2023)
-
Temporomandibular Joint Fibrocartilage Contains CD105 Positive Mouse Mesenchymal Stem/Progenitor Cells with Increased Chondrogenic Potential
Journal of Maxillofacial and Oral Surgery (2023)
-
Integrated regulation of chondrogenic differentiation in mesenchymal stem cells and differentiation of cancer cells
Cancer Cell International (2022)