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Quantitative Comparison of cis-Regulatory Element (CRE) Activities in Transgenic Drosophila melanogaster
Published on: December 19, 2011
Regulatory elements in the rat betaB2-crystallin promoter.
L Doerwald1, H Nijveen, A Civil
1Department of Biochemistry, Faculty of Science, University of Nijmegen, 6500 HB Nijmegen, The Netherlands.
Experimental Eye Research
|December 19, 2001
Summary
The transcription factor c-Maf does not appear to regulate the rat betaB2-crystallin gene promoter in lens cells, despite the presence of a Maf responsive element. Other regulatory elements likely control betaB2-crystallin gene expression in the eye lens.
Area of Science:
- Molecular and Developmental Biology
- Ophthalmology
- Gene Regulation
Background:
- The transcription factor c-Maf is proposed as a common regulator for eye lens crystallin genes.
- Maf responsive elements (MAREs) are found in crystallin gene promoters, including rat betaB2-crystallin.
- A discrepancy exists regarding c-Maf presence in rat lenses and betaB2-crystallin gene activity timing.
Purpose of the Study:
- To investigate the role of the Maf responsive element (MARE) in regulating the rat betaB2-crystallin promoter activity.
- To clarify the function of c-Maf in the context of betaB2-crystallin gene expression in rat lens fiber cells.
Main Methods:
- In vitro differentiation of rat lens fiber cells and Chinese Hamster Ovary (CHO) cells.
- Reporter gene assays to measure promoter activity.
- Deletion analysis of the betaB2-crystallin promoter, specifically targeting the MARE.
- Expression of dominant-negative c-Maf constructs.
Main Results:
- c-Maf enhanced betaB2-crystallin promoter activity in both lens fiber and CHO cells.
- Deletion of the betaB2 MARE (-143/-123) did not reduce promoter activity in lens fiber cells.
- A dominant-negative c-Maf construct failed to inhibit betaB2 promoter activity in lens fiber cells.
Conclusions:
- The betaB2 MARE is not a major regulator of the rat betaB2-crystallin promoter in lens fiber cells.
- A putative Sox binding site (-164/-159) and a positive element (-14/-7) appear to be key regulatory elements for the betaB2-crystallin promoter.
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