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Department of Pediatrics, Stanford University Medical Center, Stanford, California 94305
The GATA family of transcription
factors regulate tissue-specific patterns of gene expression during
development. We have characterized the interaction between GATA
proteins and the lactase gene promoter. Nuclear protein bound to the
lactase gene GATA region cis element (
97 to 73) was
analyzed by electrophoretic mobility shift assays (EMSA) and supershift
assays with GATA antibodies. Lactase promoter activities were assayed
in Caco-2 cells transfected with wild-type and mutated luciferase
promoter-reporter constructs and GATA-4/5/6 expression constructs. EMSA
with the GATA region probe yields a specific DNA-protein complex that
requires the GATA factor binding site WGATAR. The complex is recognized
by GATA-4- and GATA-6-specific antibodies. GATA-4/5/6 expression constructs are able to activate transcription driven by the wild-type promoter, but not by a promoter in which the GATA binding site is
mutated, in Caco-2 and nonintestinal QT6 cells. GATA factor binding to
the lactase cis element correlates with functional promoter
activation. We conclude that each of the GATA family zinc finger
proteins expressed in the intestine, GATA-4, -5, and -6, can interact
with the lactase promoter GATA element and can function to activate the
promoter in Caco-2 cells.
GATA-4; GATA-5; GATA-6; enterocyte
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