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J Biol Chem, Vol. 275, Issue 5, 3100-3106, February 4, 2000
The Pit-1 Homeodomain and -Domain Interact with Ets-1 and
Modulate Synergistic Activation of the Rat Prolactin Promoter*
Andrew P.
Bradford §¶,
Kelley S.
Brodsky ,
Scott E.
Diamond ,
Laura C.
Kuhn,
Yingmiao
Liu**, and
Arthur
Gutierrez-Hartmann§ **
From the Departments of Obstetrics and
Gynecology, § Biochemistry and Molecular Genetics, and
Medicine, and ** Program in Molecular Biology, University of
Colorado Health Sciences Center, Denver, Colorado 80262
Pit-1/GHF-1 is a pituitary-specific, POU
homeodomain transcription factor required for development of
somatotroph, lactotroph, and thyrotroph cell lineages and regulation of
the temporal and spatial expression of the growth hormone, prolactin
(PRL), and thyrotropin- genes. Synergistic interaction of Pit-1 with
a member of the Ets family of transcription factors, Ets-1, has been
shown to be an important mechanism regulating basal and Ras-induced lactotroph-specific rat (r) PRL promoter activity. Pit-1 /GHF-2, an
alternatively spliced isoform containing a 26-amino acid insert ( -domain) within its transcription-activation domain, physically interacts with Ets-1 but fails to synergize. By using a series of Pit-1
internal-deletion constructs in a transient transfection protocol to
reconstitute rPRL promoter activity in HeLa cells, we have determined
that the functional and physical interaction of Pit-1 and Ets-1 is
mediated via the POU homeodomain, which is common to both Pit-1 and
Pit-1 . Although the Pit-1 homeodomain is both necessary and
sufficient for direct binding to Ets-1 in a DNA-independent manner, an
additional interaction surface was mapped to the -domain, specific
to the Pit-1 isoform. Thus, the unique transcriptional properties of
Pit-1 and Pit-1 on the rPRL promoter may be due to the formation of
functionally distinct complexes of these two Pit-1 isoforms with
Ets-1.
*
This work was supported by National Institutes of Health
Grants DK 46868 (to A. G.-H.) and DK 53496 (to A. P. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
¶
To whom correspondence should be addressed: Dept. of
Obstetrics and Gynecology, University of Colorado Health Sciences
Center, 4200 East Ninth Ave., B-198, Denver, CO 80262. Tel.:
303-315-4146; Fax: 303-315-8889; E-mail:
Andy.Bradford@uchsc.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 2000 by the American Society for Biochemistry and Molecular Biology.
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