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(Received for publication, February 22, 1996, and in revised form, July 19, 1996)
From the Activation of the rat prolactin (rPRL) promoter
by Ras is a prototypical example of tissue-specific transcriptional
regulation in a highly differentiated cell type. Using a series of
site-specific mutations and deletions of the proximal rPRL promoter we
have mapped the major Ras/Raf response element (RRE) to a composite
Ets-1/GHF-1 binding site located between positions
Volume 271, Number 40,
Issue of October 4, 1996
pp. 24639-24648
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
,
,
,
Departments of Medicine and of Biochemistry,
Biophysics and Genetics, Program in Molecular Biology, and Colorado
Cancer Center, University of Colorado Health Sciences Center, Denver,
Colorado 80262 and the ¶ Department of Molecular Genetics, Ohio
State University, Columbus, Ohio 43210
217 and 190.
Mutation of either the Ets-1 or GHF-1 binding sites inhibits Ras and
Raf activation of the rPRL promoter, and insertion of this RRE into the
rat growth hormone promoter confers Ras responsiveness. We show that
Ets-1 is expressed in GH4 cells and, consistent with their
functional synergistic interaction, both Ets-1 and GHF-1 are able to
bind specifically to this bipartite RRE. We confirm that Ets-1 or a
related Ets factor is the nuclear target of the Ras pathway leading to
activation of the rPRL promoter and demonstrate that Elk-1 and Net do
not mediate the Ras response. Thus, the pituitary-specific POU
homeodomain transcription factor, GHF-1, serves as a cell-specific
signal integrator by functionally interacting with an Ets-1-like
factor, at uniquely juxtaposed binding sites, thereby targeting an
otherwise ubiquitous Ras signaling pathway to a select subset of
cell-specific GHF-1-dependent genes.
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