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J. Biol. Chem., Vol. 265, Issue 36, 22063-22066, 12, 1990
MS German, LG Moss and WJ Rutter
To study the regulation of insulin gene expression by physiological
regulators, primary cultures of rat islet cells were transfected with
portions of the rat insulin I gene 5'-flanking sequence linked to the
reporter gene chloramphenicol acetyltransferase (CAT). Incubation of the
cells in increasing glucose concentrations led to a parallel increase in
both CAT activity and CAT mRNA levels. Pretreatment of the cells with the
beta-cell-specific toxin streptozotocin reduced CAT activity 97%.
Beta-Cell-specific expression of CAT was also demonstrated by co-staining
the transfected cells with antisera to both CAT and insulin. Experiments
showing a reduction in the response to glucose in the presence of the
calcium channel blocker verapamil suggest that calcium plays a role in the
glucose response, possibly via regulation of factors interacting with this
limited portion of the insulin gene.
Regulation of insulin gene expression by glucose and calcium in transfected primary islet cultures
Hormone Research Institute, University of California, San Francisco 94143-0534.
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