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Endocrinology Vol. 140, No. 10 4542-4550
Copyright © 1999 by The Endocrine Society

ARTICLES

CCAAT/Enhancer-Binding Protein {alpha} Is a Physiological Regulator of Prolactin Gene Expression1

Kirsten K. Jacob2 and Frederick M. Stanley

Departments of Medicine and Pharmacology, New York University School of Medicine, New York, New York 10016

Address all correspondence and requests for reprints to: Dr. Frederick M. Stanley, Department of Medicine, TH 450, New York University Medical Center, 550 First Avenue, New York, New York 10016. E-mail: stanlf01{at}mcrcr6.med.nyu.edu

The sequence -101/-92 of the PRL promoter has been shown to be essential for both basal and hormone-increased PRL gene transcription. It is important to identify transcription factors that bind to this sequence if we are to understand the regulation of the PRL gene. Nuclear proteins, metabolically labeled with 35S were used in gel mobility shift experiments to examine which protein(s) binds to this region of the PRL promoter. An abundant 43-kDa protein binds to the PRL promoter at -106/-87. Two 43-kDa transcription factors were identified in cytosolic extracts of GH4 cells, CCAAT enhancer-binding protein {alpha} (C/EBP{alpha}) and cAMP response element-binding protein. Both of these bind to the PRL promoter, and both were present in GH4 cell nuclear extract, but only C/EBP{alpha} was definitively identified in complexes with PRL promoter DNA. Expression of C/EBP{alpha} increased basal PRL gene expression almost 6-fold, whereas expression of Chop10 that can act as an inhibitor of C/EBP{alpha} reduced the basal activity of the PRL promoter 60–75%. Mutational analysis demonstrated that the ability of C/EBP{alpha} to increase basal expression of the PRL promoter was dependent on the sequence -101/-92. These data suggest that C/EBP{alpha} is an important transcription factor that regulates PRL gene expression.




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