PMID:20530484

Citation	Masuda, T and Esumi, N (2010) SOX9, through interaction with microphthalmia-associated transcription factor (MITF) and OTX2, regulates BEST1 expression in the retinal pigment epithelium. J. Biol. Chem. 285:26933-44
Abstract	BEST1 is highly and preferentially expressed in the retinal pigment epithelium (RPE) and causes Best macular dystrophy when mutated. We previously demonstrated that the human BEST1 upstream region -154 to +38 bp is sufficient to direct expression in the RPE of transgenic mice, and microphthalmia-associated transcription factor (MITF) and OTX2 regulate this BEST1 promoter. However, a number of questions remained. Here, we show that yeast one-hybrid screen with bait corresponding to BEST1 -120 to -88 bp identified the SOX-E factors, SOX8, SOX9, and SOX10. A paired SOX site was found in this bait, and mutation of either of the paired sites significantly decreased BEST1 promoter activity in RPE primary cultures. Among the SOX-E genes, SOX9 is highly and preferentially expressed in the RPE, and chromatin immunoprecipitation with fresh RPE cells revealed binding of SOX9, but not SOX10, to the BEST1 region where the paired SOX site is located. BEST1 promoter activity was increased by SOX9 overexpression and decreased by siRNA-mediated SOX9 knockdown. Importantly, SOX9 physically interacted with MITF and OTX2 and orchestrated synergistic activation of the BEST1 promoter with the paired SOX site playing essential roles. A combination of the expression patterns of SOX9, MITF, and OTX2 yielded tissue distribution remarkably similar to that of BEST1. Lastly, the BEST1 promoter was also active in Sertoli cells of the testis in transgenic mice where SOX9 is highly expressed. These results define SOX9 as a key regulator of BEST1 expression and demonstrate for the first time its functional role in the RPE.
Links	PubMed PMC2930693 Online version:10.1074/jbc.M110.130294
Keywords	Animals; Cell Line, Tumor; Chloride Channels/biosynthesis; Chloride Channels/genetics; Eye Proteins/biosynthesis; Eye Proteins/genetics; Gene Expression Regulation/physiology; Humans; Ion Channels; Male; Mice; Mice, Transgenic; Microphthalmia-Associated Transcription Factor/genetics; Microphthalmia-Associated Transcription Factor/metabolism; Otx Transcription Factors/genetics; Otx Transcription Factors/metabolism; Response Elements/physiology; Retinal Pigment Epithelium/cytology; Retinal Pigment Epithelium/metabolism; SOX9 Transcription Factor/genetics; SOX9 Transcription Factor/metabolism; Sertoli Cells/cytology; Sertoli Cells/metabolism; Swine
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
PIG:SOX9	enables	GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific	ECO:0000315: mutant phenotype evidence used in manual assertion		F	occurs_at:(SO:0001952)	Seeded From UniProt	complete
PIG:SOX9	involved_in	GO:0045944: positive regulation of transcription by RNA polymerase II	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:MITF	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P48436	F		Seeded From UniProt	complete
HUMAN:MITF	enables	GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific	ECO:0000314: direct assay evidence used in manual assertion		F	occurs_at:(SO:0001952)	Seeded From UniProt	complete
HUMAN:OTX2	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P48436	F		Seeded From UniProt	complete
HUMAN:OTX2	enables	GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific	ECO:0000314: direct assay evidence used in manual assertion		F	occurs_at:(SO:0001952)	Seeded From UniProt	complete
HUMAN:OTX2	involved_in	GO:0065003: protein-containing complex assembly	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:OTX2	part_of	GO:0032991: protein-containing complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:OTX2	involved_in	GO:0045944: positive regulation of transcription by RNA polymerase II	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:SOX9	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P32243	F		Seeded From UniProt	complete
HUMAN:SOX9	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:O75030	F		Seeded From UniProt	complete
HUMAN:SOX9	enables	GO:0000978: RNA polymerase II cis-regulatory region sequence-specific DNA binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:SOX9	enables	GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific	ECO:0000314: direct assay evidence used in manual assertion		F	occurs_at:(SO:0001952)	Seeded From UniProt	complete
HUMAN:SOX9	enables	GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific	ECO:0000314: direct assay evidence used in manual assertion		F	occurs_at:(SO:0001952)	Seeded From UniProt	complete
HUMAN:SOX9	involved_in	GO:0065003: protein-containing complex assembly	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:SOX9	enables	GO:0000978: RNA polymerase II proximal promoter sequence-specific DNA binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
HUMAN:SOX9	part_of	GO:0032991: protein-containing complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:SOX9	involved_in	GO:0045944: positive regulation of transcription by RNA polymerase II	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:MITF	part_of	GO:0032991: protein-containing complex	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:MITF	involved_in	GO:0065003: protein-containing complex assembly	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:MITF	enables	GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific	ECO:0000314: direct assay evidence used in manual assertion		F	occurs_at:(SO:0001952)	Seeded From UniProt	complete
HUMAN:MITF	involved_in	GO:0045944: positive regulation of transcription by RNA polymerase II	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete

References

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