| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Submitted on July 28, 2004
Accepted on January 25, 2005
Division of Biomedical Sciences, University of California, Riverside, 900 University Avenue, Riverside, CA, 92521 -0121; Mammary Biology and Tumorigenesis Laboratory, National Cancer Institute, Bethesda, MD 20892; Oncology Research Inst., Clemson University, Greenville, SC 29605
* To whom correspondence should be addressed. E-mail: Ameae.Walker{at}ucr.edu.
We have used bioluminescence resonance energy transfer (BRET) to examine the interaction between human PRLs and the long (LF) and two short isoforms (SF1a and SF1b) of the human PRL receptor in living cells. cDNA sequences encoding the LF, SF1a and SF1b were subcloned into codon-humanized vectors containing cDNAs for either luciferase (Rluc) or a green fluorescent protein (GFP2) with a 12- or 13- amino acid linker connecting the parts of the fusion proteins. Transfection into HEK 293 cells demonstrated maintained function of Rluc and GFP2 when linked to the receptors and confocal microscopy demonstrated the localization of tagged receptors in the plasma membrane by 48 h posttransfection. All three tagged receptors transduced a signal, with the LF and SF1a stimulating, and SF1b inhibiting, promoter activity of a 
2.4kb 
-casein-luc construct. Both unmodified (U) PRL and the molecular mimic of phosphorylated PRL, S179D PRL, induced BRET with all combinations of long and short receptor isoforms except SF1a plus SF1b. No BRET was observed with the site two-inactive mutant, G129R PRL. This is the first demonstration, (1) that species homologous PRL promotes both homo- and hetero-interaction of most long and short PRLR pairs in living cells, (2) that both U-PRL and S179D PRL are active in this regard and (3) that there is some aspect of SF1a-SF1b structure that prevents this particular hetero-receptor pairing. In addition, we conclude that preferential pairing of different receptor isoforms is not the explanation for the different signaling initiated by U-PRL and S179D PRL.
This article has been cited by other articles:
 |
|
 |
|
  A. Bachelot and N. Binart Reproductive role of prolactin Reproduction, February 1, 2007; 133(2): 361 - 369. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. L. Gadd and C. V. Clevenger Ligand-Independent Dimerization of the Human Prolactin Receptor Isoforms: Functional Implications Mol. Endocrinol., November 1, 2006; 20(11): 2734 - 2746. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. K. Ueda, H.-L. Lo, P. Bartolini, and A. M. Walker S179D Prolactin Primarily Uses the Extrinsic Pathway and Mitogen-Activated Protein Kinase Signaling to Induce Apoptosis in Human Endothelial Cells Endocrinology, October 1, 2006; 147(10): 4627 - 4637. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Ueda, U. Ozerdem, Y.-H. Chen, M. Yao, K. T. Huang, H. Sun, M. Martins-Green, P. Bartolini, and A. M Walker A molecular mimic demonstrates that phosphorylated human prolactin is a potent anti-angiogenic hormone. Endocr. Relat. Cancer, March 1, 2006; 13(1): 95 - 111. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. F. Langenheim, D. Tan, A. M. Walker, and W. Y. Chen Two Wrongs Can Make a Right: Dimers of Prolactin and Growth Hormone Receptor Antagonists Behave as Agonists Mol. Endocrinol., March 1, 2006; 20(3): 661 - 674. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Wu, E. Ginsburg, B. K. Vonderhaar, and A. M. Walker S179D Prolactin Increases Vitamin D Receptor and p21 through Up-regulation of Short 1b Prolactin Receptor in Human Prostate Cancer Cells Cancer Res., August 15, 2005; 65(16): 7509 - 7515. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| Endocrinology | Endocrine Reviews | J. Clin. End. & Metab. |
| Molecular Endocrinology | Recent Prog. Horm. Res. | All Endocrine Journals |
