Receptor activator of NF-kappaB ligand induction via Jak2 and Stat5a in mammary epithelial cells.
Prolactin (PRL) is the primary hormone that, in conjunction with local factors, leads to lobuloalveolar development during pregnancy. Recently, receptor activator of NF-kappaB 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-kappaB) 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 gamma-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.
Srivastava
Sunil
S
Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, Ohio 45267-0576, USA.
Matsuda
Manabu
M
Hou
Zhaoyuan
Z
Bailey
Jason P
JP
Kitazawa
Riko
R
Herbst
Matthew P
MP
Horseman
Nelson D
ND
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D005799:Genes, Dominant; D006023:Glycoproteins / Q000096:biosynthesis; D007371:Interferon-gamma / Q000378:metabolism; D053614:Janus Kinase 2; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008894:Milk Proteins; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D053244:Osteoprotegerin; D010957:Plasmids / Q000378:metabolism; D011388:Prolactin / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D053245:RANK Ligand; D012333:RNA, Messenger / Q000378:metabolism; D053246:Receptor Activator of Nuclear Factor-kappa B; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis; D018124:Receptors, Tumor Necrosis Factor; D050799:STAT5 Transcription Factor; D013997:Time Factors; D015534:Trans-Activators / Q000737:chemistry*; D014162:Transfection
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D008024:Ligands; D008562:Membrane Glycoproteins; D008894:Milk Proteins; D053244:Osteoprotegerin; D011518:Proto-Oncogene Proteins; D053245:RANK Ligand; D012333:RNA, Messenger; D053246:Receptor Activator of Nuclear Factor-kappa B; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; D050799:STAT5 Transcription Factor; C494093:Stat5a protein, mouse; C505687:Tnfrsf11a protein, mouse; C506031:Tnfrsf11b protein, mouse; C505689:Tnfsf11 protein, mouse; D015534:Trans-Activators; D007371:Interferon-gamma; D011388:Prolactin; D008156:Luciferases; D011505:Protein-Tyrosine Kinases; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2