The dimerization domain of SOX9 is required for transcription activation of a chondrocyte-specific chromatin DNA template.
Mutations in SOX9, a gene essential for chondrocyte differentiation cause the human disease campomelic dysplasia (CD). To understand how SOX9 activates transcription, we characterized the DNA binding and cell-free transcription ability of wild-type SOX9 and a dimerization domain SOX9 mutant. Whereas formation of monomeric mutant SOX9-DNA complex increased linearly with increasing SOX9 concentrations, formation of a wild-type SOX9-DNA dimeric complex increased more slowly suggesting a more sigmoidal-type progression. Stability of SOX9-DNA complexes, however, was unaffected by the dimerization mutation. Both wild-type and mutant SOX9 activated transcription of a naked Col2a1 DNA template. However, after nucleosomal assembly, only wild-type and not the mutant was able to remodel chromatin and activate transcription of this template. Using a cell line, in which the Col2a1 vector was stably integrated, no differences were seen in the interactions of wild-type and mutant SOX9 with the chromatin of the Col2a1 vector using ChIP. However, the mutant was unable to activate transcription in agreement with in vitro results. We hypothesize that the SOX9 dimerization domain is necessary to remodel the Col2a1 chromatin in order to allow transcription to take place. These results further clarify the mechanism that accounts for CD in patients harboring SOX9 dimerization domain mutations.
Coustry
Françoise
F
Department of Genetics, The University of Texas, MD Anderson Cancer Center, Houston, TX 77030, USA.
Oh
Chun-do
CD
Hattori
Takako
T
Maity
Sankar N
SN
de Crombrugghe
Benoit
B
Yasuda
Hideyo
H
D000818:Animals; D002460:Cell Line; D019902:Chondrocytes / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D024043:Collagen Type II / Q000235:genetics; D004247:DNA / Q000378:metabolism; D019281:Dimerization; D004742:Enhancer Elements, Genetic; D006801:Humans; D009154:Mutation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015533:Transcriptional Activation
D002843:Chromatin; D024043:Collagen Type II; D011994:Recombinant Proteins; D055758:SOX9 Transcription Factor; D004247:DNA