Gonadotropin-releasing hormone-mediated phosphorylation of estrogen receptor-alpha contributes to fosB expression in mouse gonadotrophs.
Estrogen receptors (ERs) are activated by their ligands as well as signaling pathways that alter ER phosphorylation in response to peptide hormones and growth factors. In pituitary gonadotrophs, GnRHs act via the type I GnRH receptor (GnRHR). Both GnRH subtypes (GnRH-I and -II) activate an estrogen response element (ERE)-driven luciferase reporter gene in LbetaT2 mouse pituitary cells, and GnRH-I is most potent in this regard. Moreover, antide (a GnRH antagonist) and a GnRHR small interfering RNA (siRNA) abrogate this effect, whereas an ERalpha antagonist (ICI 182,780) does not. The ERalpha in LbetaT2 cells is phosphorylated at Ser(118) in the nucleus and at Ser(167) in both nucleus and cytoplasm after GnRH treatments and coincided with increased ERalpha binding to its coactivator, the p300/cAMP response element-binding protein-associated factor (PCAF). Moreover, siRNA-mediated knockdown of PCAF levels attenuated GnRH-induced ERE-luciferase transactivation in these cells. Most importantly, both GnRH subtypes robustly up-regulated expression of the immediate early response gene, fosB, whereas cotreatment with ERalpha siRNA or PCAF siRNA attenuated this effect. This appears to occur at the transcriptional level because corecruitment of ERalpha and PCAF to an ERE within the endogenous fosB promoter was increased by GnRH treatments, as shown by chromatin immunoprecipitation assays. These data demonstrate that GnRH-mediated phosphorylation of ERalpha in mouse LbetaT2 pituitary cells results in its rapid association with PCAF and the transcriptional activation of fosB, and we demonstrate that this in turn likely activates other genes in pituitary cells including the FSH beta-subunit gene.
Chen
Junling
J
Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada.
An
Beum-Soo
BS
Cheng
Linan
L
Hammond
Geoffrey L
GL
Leung
Peter C K
PC
D000818:Animals; D002460:Cell Line; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D037201:Follicle Stimulating Hormone, beta Subunit / Q000378:metabolism; D052681:Gonadotrophs / Q000378:metabolism*; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D008156:Luciferases; D051379:Mice; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism*; D020218:Response Elements; D050880:p300-CBP Transcription Factors / Q000378:metabolism*; D000099224:p300-CBP-Associated Factor
D047628:Estrogen Receptor alpha; D037201:Follicle Stimulating Hormone, beta Subunit; D007987:Gonadotropin-Releasing Hormone; D008156:Luciferases; D010770:Phosphotransferases; D016760:Proto-Oncogene Proteins c-fos; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C483819:Fosb protein, mouse