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J. Biol. Chem., Vol. 278, Issue 46, 46171-46178, November 14, 2003
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Receptor Activator of NF-
B Ligand Induction via Jak2 and Stat5a in Mammary Epithelial Cells*
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From the
Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, Ohio 45267-0576, theDepartment of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-003, Japan, and the ¶Department of Pathology, Kobe University, Kobe 650-0017, Japan
Prolactin (PRL) is the primary hormone that, in conjunction with local factors, leads to lobuloalveolar development during pregnancy. Recently, receptor activator of NF-
B ligand (RANKL) has been identified as one of the effector molecules essential for lobuloalveolar development. The molecular mechanisms by which PRL may induce RANKL expression have not been carefully examined. Here we report that RANKL expression in the mammary gland is developmentally regulated and dependent on PRL and progesterone, whereas its receptor RANK (receptor activator of NF-B) and decoy receptor osteoprotegerin (OPG) are constitutively expressed at all stages in both normal (PRL+/-) and prolactin knockout (PRL-/-) mice. In vitro, PRL markedly increased RANKL expression in primary mammary epithelial cells and RANKL-luciferase reporter activity in CHOD6 cells, which constitutively express the PRL receptor. We identified a 
-interferon activation sequence (GAS) in the region between residues -965 to -725 of the RANKL promoter, which conferred a PRL response. Using dominant negative mutants of recombinant Jak2 and Stat5 in CHOD6 cells, and by reconstituting the Jak2/Stat5 pathway in COS7 cells, we determined that Jak2 and Stat5a are essential for the PRL-induced RANKL expression in mammary gland.
Received for publication, August 4, 2003
* This work was supported by National Institutes of Health Grant DK-53124 and grants from the Shriners Hospital for Children (to N. D. H.) and National Osteoporosis Foundation grant (to S. S.). The costs of publication of this article were defrayed in part by the payment of page charges. This 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 Molecular and Cellular Physiology, 231 Albert Sabin Way, Cincinnati, OH 45267-0576. Tel.: 513-558-3019; Fax: 513-558-5738; E-mail: nelson.horseman{at}uc.edu.
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