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Investigative Ophthalmology & Visual Science, Vol 38, 951-959, Copyright © 1997 by Association for Research in Vision and Ophthalmology

ARTICLES AND REPORTS

A complex enhancer of the chicken beta A3/A1-crystallin gene depends on an AP-1-CRE element for activity

JB McDermott, A Cvekl and J Piatigorsky
Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892- 2730, USA.

PURPOSE: To define transcriptional regulatory elements of the chicken beta A3/A1-crystallin gene. METHODS: Reporter genes were made with fragments of the chicken beta A3/A1-crystallin gene fused to the bacterial gene encoding chloramphenicol acetyltransferase (CAT). The reporter plasmids were transfected into primary cultures of chicken- patched lens epithelium or fibroblast cells, and the CAT activity of cellular extracts was measured. The binding of lens nuclear proteins to beta A3/A1 sequences was tested in electrophoretic mobility shift assays. RESULTS: Sequences from -287 to -254 bp relative to the transcriptional start site function as an enhancer in transfected lens and nonlens cells. The length of a T-rich sequence downstream of the enhancer influences its activity. Minimal enhancer activity depends on sequences between -270 and -254 bp, and full activity requires additional upstream sequences. The minimal enhancer includes a consensus sequence (TGAGTCA) for basic region-leucine zipper (bZIP) proteins of the AP-1-CREB superfamily. Lens nuclear proteins bind the enhancer sequences to form several specific complexes, some of which are related antigenically to members of the AP-1 and CREB families of proteins. CONCLUSIONS: An enhancer of the chicken beta A3/A1-crystallin gene between -287 and -254 bp functions in both lens and nonlens cells and binds multiple nuclear proteins. Temporal and spatial regulation of beta A3/A1 expression in the lens may be regulated by the enhancer.


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Copyright © 1997 by the Association for Research in Vision and Ophthalmology