Nucleic Acids Research, Vol 24, Issue 6 1065-1072, Copyright © 1996 by Oxford University Press
SD Willis and MA Seyfred
The location of target DNA sequences within chromatin may affect the
ability of trans-acting factors to bind cis-elements and regulate gene
transcription. To examine the effect of chromatin structure on the ability
of the estrogen-estrogen receptor complex (E2R) to bind its respective DNA
binding element within the rat prolactin (rPRL) gene and modulate rPRL gene
expression, we have developed cell lines derived from the rPRL-expressing
(rPRL+) rat pituitary cell line GH3 and the rPRL-non- expressing (rPRL-)
rat embryo fibroblast cell line Rat1. These cell lines contain
mini-chromosomes composed of the 5' upstream regulatory region of the rPRL
gene driving expression of a reporter gene, Tn5, within a bovine
papillomavirus (BPV) vector. The rPRL-Tn5 gene retains the characteristics
of cell-specific expression and estrogen inducibility of transcription
displayed by the endogenous rPRL gene. The distal enhancer region, which
contains an estrogen response element, was found to exist in a
nucleosome-free region in pituitary- derived cells even in the absence of
estrogen. In contrast, the rPRL distal enhancer in fibroblast cells was
found to be randomly packaged into nucleosomes. These results indicate that
DNA sequence is not sufficient to position nucleosomes in the rPRL gene.
Rather, it suggests that cell-specific factors are present in pituitary
cells that modify the chromatin structure of the distal enhancer which
allow E2R to bind to its response element.
ARTICLES
Pituitary-specific chromatin structure of the rat prolactin distal enhancer element
Department of Molecular Biology, Vanderbilt University, Nashville, TN 37235 USA.
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