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Papers In Press, published online ahead of print August 27, 2002
Department of Basic Allied Medicine, Kobe University School of Medicine, Kobe 654-0142
Corresponding Author: okimura{at}ams.kobe-u.ac.jp
Pit-1 stimulates the expression of growth hormone, prolactin, and thyrotropin b 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 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 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.
J. Biol. Chem, 10.1074/jbc.M202991200
Submitted on March 27, 2002
Revised on August 27, 2002
Accepted on August 27, 2002
Novel function of transactivation domain of the pituitary specific transcription factor, Pit-1
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