This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hou, Z. Articles by Horseman, N. D. Search for Related Content PubMed PubMed Citation Articles by Hou, Z. Articles by Horseman, N. D.

Two Tandemly Linked Interferon--Activated Sequence Elements in the Promoter of Glycosylation-Dependent Cell Adhesion Molecule 1 Gene Synergistically Respond to Prolactin in Mouse Mammary Epithelial Cells

Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, Ohio 45267

Address all correspondence and requests for reprints to: Nelson D. Horseman, Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Albert Sabin Way, Cincinnati, Ohio 45267-0576. E-mail: nelson.horseman{at}uc.edu.

Previously, we reported that glycosylation-dependent cell adhesion molecule 1 (GlyCAM 1) was a novel target for prolactin (PRL) in the mouse mammary gland. However, the signaling pathway by which PRL regulates GlyCAM 1 expression has not been specified. In the present study, we showed that PRL induced GlyCAM 1 expression in primary mammary epithelial cells of mice through the Janus kinase 2/signal transducer and activator of transcription 5 (Stat5) pathway. Deletion and site-directed mutagenesis analyses of the GlyCAM 1 promoter demonstrated that the two tandemly linked Stat5 binding sites [interferon--activated sequence 1 and -2 (GAS1 and GAS2)] in the proximal promoter region were crucial and synergistically responded to PRL. GAS2, a consensus GAS site, was essential and, by itself, weakly responded to PRL, whereas GAS1, a nonconsensus site, failed to respond to PRL but was indispensable for the maximal activity of the GlyCAM 1 promoter. Gel shift assays showed that probe containing GAS1 and GAS2 bound two Stat5 complexes, which represent Stat5 dimer and tetramer, respectively, while GAS2, by itself, bound Stat5 as a dimer only, and GAS1 showed no apparent binding activity. Interruption of tetramer formation by mutation of a tryptophan to alanine (W37A), and a leucine to serine (L83S) in the N terminus of Stat5A attenuated the synergistic effect between the two tandemly linked GAS sites. Overexpression of W37A and L83S mutants in primary mammary epithelial cells suppressed endogenous GlyCAM 1 expression.

This article has been cited by other articles:

				Z. Hou, H. Peng, K. Ayyanathan, K.-P. Yan, E. M. Langer, G. D. Longmore, and F. J. Rauscher III The LIM Protein AJUBA Recruits Protein Arginine Methyltransferase 5 To Mediate SNAIL-Dependent Transcriptional Repression Mol. Cell. Biol., May 15, 2008; 28(10): 3198 - 3207. [Abstract] [Full Text] [PDF]

				S. Srivastava, M. Matsuda, Z. Hou, J. P. Bailey, R. Kitazawa, M. P. Herbst, and N. D. Horseman Receptor Activator of NF-{kappa}B Ligand Induction via Jak2 and Stat5a in Mammary Epithelial Cells J. Biol. Chem., November 14, 2003; 278(46): 46171 - 46178. [Abstract] [Full Text] [PDF]