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J. Biol. Chem., Vol. 277, Issue 47, 45141-45148, November 22, 2002
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From the Division of Endocrinology/Metabolism, Neurology and
Hematology/Oncology, Pit-1 stimulates the expression of growth
hormone, prolactin, and thyrotropin
Novel Function of the Transactivation Domain of a
Pituitary-specific Transcription Factor, Pit-1*
,
,,,,, and
Department of Clinical Molecular
Medicine, and § Department of Basic Allied Medicine,
Department of Brain Sciences, Kobe University Graduate
School of Medicine, Kobe University School of Medicine, 7-10-2, Tomogaoka, Suma-ku, Kobe 654-0142, Japan, and the ** College
of Nursing Art and Culture, Hyogo 655-0048, Japan
subunit genes.
Consequently, abnormality of the Pit-1 gene results
in combined pituitary hormone deficiency (CPHD). In this study, we
analyzed the function of Pit-1 with a mutation (proline to leucine at
codon 24) in the transactivation domain, P24L, which has a normal POU
domain important for binding to DNA, because this mutation had been
reported in a patient with CPHD. We found that codon 24 proline in the
transactivation domain as well as the POU domain of Pit-1 was crucial
to recruit coactivator CREB-binding protein (CBP) in the cultured
cells. P24L completely lost the responsiveness to cAMP to stimulate the
expression of the Pit-1-targeted genes. Furthermore, CBP and Pit-1, but
not P24L, markedly enhanced the expression of the Pit-1-targeted gene to cAMP, and adenovirus E1a that binds to CBP and abrogates its function blocked the induction by cAMP of Pit-1-stimulated gene transcription in the pituitary-derived GH3 cells. These results suggest
that CBP and proline at codon 24 in the transactivation domain of Pit-1
are important for the cAMP-induced activation of Pit-1-targeted genes.
However, P24L maintained basal transcriptional activity, suggesting
that CBP is unlikely to be an essential coactivator for
Pit-1.
*
This work was supported in part by a grant-in-aid for
Scientific Research from the Japanese Ministry of Education, Science, Sports, and Culture, and grants from the Japanese Ministry of Health,
Labor and Welfare and from Growth Science Research Foundation.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.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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