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PMID:11139622

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Citation

Lengler, J, Krausz, E, Tomarev, S, Prescott, A, Quinlan, RA and Graw, J (2001) Antagonistic action of Six3 and Prox1 at the gamma-crystallin promoter. Nucleic Acids Res. 29:515-26

Abstract

Gamma-crystallin genes are specifically expressed in the eye lens. Their promoters constitute excellent models to analyse tissue-specific gene expression. We investigated murine CRYGE/f promoters of different length in lens epithelial cell lines. The most active fragment extends from position -219 to +37. Computer analysis predicts homeodomain and paired-domain binding sites for all rodent CRYGD/e/f core promoters. As examples, we analysed the effects of Prox1 and Six3, which are considered important transcription factors involved in lens development. Because of endogenous Prox1 expression in N/N1003A cells, a weak stimulation of CRYGE/f promoter activity was found for PROX1. In contrast, PROX1 stimulated the CRYGF promoter 10-fold in CD5A cells without endogenous PROX1. In both cell lines Six3 repressed the CRYGF promoter to 10% of its basal activity. Our cell transfection experiments indicated that CRYG expression increases as Six3 expression decreases. Prox1 and Six3 act antagonistically on regulation of the CRYGD/e/f promoters. Functional assays using randomly mutated gammaF-crystallin promoter fragments define a Six3-responsive element between -101 and -123 and a Prox1-responsive element between -151 and -174. Since Prox1 and Six3 are present at the beginning of lens development, expression of CRYGD/e/f is predicted to remain low at this time. It increases as Six3 expression decreases during ongoing lens development.

Links

PubMed PMC29665

Keywords

3T3 Cells; Animals; Base Sequence; Binding Sites/genetics; Cell Line; Consensus Sequence/genetics; Crystallins/antagonists & inhibitors; Crystallins/genetics; Eye Proteins/genetics; Eye Proteins/physiology; Gene Expression Regulation/genetics; Homeodomain Proteins/biosynthesis; Homeodomain Proteins/genetics; Homeodomain Proteins/physiology; Humans; Mice; Molecular Sequence Data; Nerve Tissue Proteins/biosynthesis; Nerve Tissue Proteins/genetics; Nerve Tissue Proteins/physiology; Paired Box Transcription Factors; Promoter Regions, Genetic/genetics; Rabbits; Rats; Repressor Proteins/genetics; Sequence Alignment; Tumor Suppressor Proteins

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