Nature Genetics
13, 230 - 232 (1996)
doi:10.1038/ng0696-230
Sex reversal by loss of the C−terminal transactivation domain of human SOX9Peter Südbeck1, M. Lienhard Schmitz2, Patrick A. Baeuerle2, 3
& Gerd Scherer1
1Institute of Human Genetics, University of Freiburg, Breisacherstr. 33, D-79106 Freiburg i. Br., Germany Institute of Biochemistry, University of Freiburg,Breisacherstr.33,D-79104 Freiburg i.Br., Germany
2Institute of Biochemistry, University of Freiburg, Hermann-Herder-Str. 7,D-79104 Freiburg i. Br., Germany
3Current address:Tularik Inc., 270 E. Grand Ave., San Francisco, California 94080, USA Correspondence should be addressed to G.S. Haploinsufficiency for SOX9 has recently been identified as the cause for both campomelic dysplasia (CD), a human skeletal malformation syndrome, and the associated autosomal XY sex reversal1,2. SOX9 contains a putative DNA-binding motif known as the high-mobility group (HMG) domain characterizing a whole class of transcription factors3. We show in cell transfection experiments that SOX9 can trans-activate transcription from a reporter plasmid through the motif AACAAAG, a sequence recognized by other HMG domain transcription factors3. By fusing all or part of SOX9 to the DNA-binding domain of yeast GAL4, the transactivating function was mapped to a transcription activation (TA) domain at the C terminus of SOX9. This non-acidic TA domain is evolutionary conserved and rich in proline, glutamine and serine. With one exception4, all SOX9 nonsense and frame shift mutations described so far in CD/sex reversal patients1,2,4 lead to truncation of the TA domain, suggesting that impairment of gonadal and skeletal development in these cases results, at least in part, from loss of transactivation of genes downstream of SOX9.
REFERENCES
- 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).
- Foster, J.W. et al. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. 372, 525−530 (1994).
- Grosschedl, R., Giese, K. & Pagel, J. HMG domain proteins: architectural elements in the assembly of nucleoprotein structures.Trends Genet. 10, 94−100 (1994).
- Kwok, C. et al. Mutations in SOX9, the gene responsible for campomelic dysplasia and autosomal sex reversal. Am. J. Hum. Genet. 57, 1028−1036 (1995).
- van de Wetering, M., Oosterwegel, M., Dooijes, D. & Clevers, H. Identification and cloning of TCF-1, a T lymphocyte-specific transcription factor containing a sequence-specific HMG box. EMBO J. 10, 123−132 (1991).
- Harley, V.R. et al. DNA binding activity of recombinant SRY from normal mates and XY females. Science 255, 453−456 (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).
- Wright, E. et al. The Sry-related gene Sox9 is expressed during chondrogenesis in mouse embryos. Nature Genet 9, 15−20 (1995).
- Tjian, R. & Maniatis, T. Transcriptional activation: a complex puzzle with few easy pieces. Cell 77, 5−8 (1994).
- Chi, T., Lieberman, P., Ellwood, K. & Carey, M. A general mechanism for transcriptional synergy by eukaryotic activators. Nature 377, 254−257 (1995).
- Gerber, H.-P. et al.Transcriptional activation modulated by homopolymeric glutamine and proline stretches. Science 263, 808−811 (1994).
- Bours, V. et al.The oncoprotein Bcl-3 directly transactivates through
 B motifs via association with DNA-binding p50B homodimers. Cell 72, 729−739 (1993).
- Glaser, T. et al. PAX6 gene dosage effect in a family with congenital cataracts, aniridia, anophthalmia and central nervous system defects. Nature Genet. 7, 463−471 (1994).
- Truant, R., Xiao, H., Ingles, C.J. & Greenblatt, J. Direct interaction between the transcriptional activation domain of human p53 and the TATA box-binding protein. J. Biol. Chem. 268, 2284−2287 (1993).
- Clark, H.M. et al.Mutations in the coding region of c-myc in AIDS-associated and other aggressive lymphomas. Cancer Res. 54, 3383−3386 (1994).
- Baniahmad, A., Köhne, A.C. & Renkawitz, R. A transferable silencing domain is present in the thyroid hormone receptor, in the v-erbA oncogene product and in the retinoic acid receptor. EMBO J. 11, 1015−1023 (1992).
- Schmitz, M.L., dos Santos Silva, M.A. & Baeuerle, P.A. Transactivation domain 2 (TA2) of p65NF-B. J. Biol. Chem. 270, 15576−15584 (1995).
- Giese, K., Amsterdam, A. & Grosschedl, R. DNA-binding properties of the HMG domain of the lymphoid-specific transcriptional regulator LEF-1. Genes Dev. 5, 2567−2578 (1991).
- Ferrari, S. et al. SRY, like HMG1, recognizes sharp angles in DNA. EMBO J. 11, 4497−4506 (1992).
- Wigler, M., Pellicer, A., Silverstein, S. & Axel, R. Biochemical transfer of single-copy eucaryotic genes using total cellar DNA as donor. Cell 14, 725−731 (1978).
- Schreiber, E., Matthias, P., Mülter, M.M. & Schaffner, W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucl. Acids Res. 17, 6419 (1989).
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