|
|
 Previous Article | Table of Contents | Next Article 
Blood, 15 April 2001, Vol. 97, No. 8, pp. 2230-2237
HEMATOPOIESIS
A common epitope is shared by activated signal transducer and
activator of transcription-5 (STAT5) and the phosphorylated
erythropoietin receptor: implications for the docking model of STAT
activation
Dwayne L. Barber,
Bryan K. Beattie,
Jacqueline M. Mason,
Melody H.-H. Nguyen,
Monique Yoakim,
Benjamin G. Neel,
Alan D. D'Andrea, and
David A. Frank
From the Division of Cellular and Molecular Biology,
Ontario Cancer Institute; Department of Laboratory Medicine and
Pathobiology, Toronto General Hospital; Departments of Medical
Biophysics and Laboratory Medicine and Pathobiology, University of
Toronto; Toronto, ON; Cancer Biology Program and Division of
Hematology-Oncology, Department of Medicine, Beth Israel Deaconess
Medical Center; and Departments of Pediatric Oncology and Adult
Oncology, Dana-Farber Cancer Institute; Boston, MA.
Erythropoietin (EPO) specifically activates the Janus kinase JAK2
and the transcription factor signal transducer and activator of
transcription-5 (STAT5). All members of the STAT family are tyrosine
phosphorylated in response to cytokine stimulation at a conserved
carboxy-terminal tyrosine, Y694, in the case of STAT5. To determine
structural features important for STAT signaling, we generated an
activation-specific STAT5 antibody using a phosphopeptide containing
amino acids 687 to 698 of STAT5 as antigen. This antibody specifically recognizes tyrosine- phosphorylated STAT5 but not nonphosphorylated STAT5. In immunoprecipitation reactions from cell
lines and primary erythroblasts, 2 distinct polyclonal
activation-specific STAT5 antibodies selectively immunoprecipitate the
tyrosine phosphorylated EPO receptor (EPO-R) in addition to STAT5 under
native and denaturing conditions. We propose that the
activation-specific STAT5 antibody recognizes the 2 substrates to which
the STAT5 SH2 domain interacts, namely, the tyrosine- phosphorylated
EPO-R and STAT5 itself. Several studies have implicated EPO-R Y343,
Y401, Y431, and Y479 in the recruitment of STAT5. Using a series of
EPO-R tyrosine mutants expressed in Ba/F3 cells, we have shown that the
activation-specific STAT5 antibody immunoprecipitates an EPO-R
containing only 2 tyrosines at positions 343 and 401, confirming the
importance of these tyrosines in STAT5 recruitment. These data uncover
a novel aspect of STAT SH2 domain recognition and demonstrate the
utility of activation-specific antibodies for examining the specificity
of STAT-cytokine receptor interactions.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
O. Ravid, I. Shams, N. Ben Califa, E. Nevo, A. Avivi, and D. Neumann
An extracellular region of the erythropoietin receptor of the subterranean blind mole rat Spalax enhances receptor maturation
PNAS,
September 4, 2007;
104(36):
14360 - 14365.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Sathyanarayana, M. P. Menon, O. Bogacheva, O. Bogachev, K. Niss, W. S. Kapelle, E. Houde, J. Fang, and D. M. Wojchowski
Erythropoietin modulation of podocalyxin and a proposed erythroblast niche
Blood,
July 15, 2007;
110(2):
509 - 518.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. V. Alvarez, H. Greulich, W. R. Sellers, M. Meyerson, and D. A. Frank
Signal transducer and activator of transcription 3 is required for the oncogenic effects of non-small-cell lung cancer-associated mutations of the epidermal growth factor receptor.
Cancer Res.,
March 15, 2006;
66(6):
3162 - 3168.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Montoye, I. Lemmens, D. Catteeuw, S. Eyckerman, and J. Tavernier
A systematic scan of interactions with tyrosine motifs in the erythropoietin receptor using a mammalian 2-hybrid approach
Blood,
June 1, 2005;
105(11):
4264 - 4271.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Martens, G. Uzan, M. Wery, R. Hooghe, E. L. Hooghe-Peters, and A. Gertler
Suppressor of Cytokine Signaling 7 Inhibits Prolactin, Growth Hormone, and Leptin Signaling by Interacting with STAT5 or STAT3 and Attenuating Their Nuclear Translocation
J. Biol. Chem.,
April 8, 2005;
280(14):
13817 - 13823.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Halupa, M. L. Bailey, K. Huang, N. N. Iscove, D. E. Levy, and D. L. Barber
A novel role for STAT1 in regulating murine erythropoiesis: deletion of STAT1 results in overall reduction of erythroid progenitors and alters their distribution
Blood,
January 15, 2005;
105(2):
552 - 561.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Tong, X. Chu, J. Y. Cheung, K. Conrad, R. Stahl, D. L. Barber, G. Mignery, and B. A. Miller
Erythropoietin-modulated calcium influx through TRPC2 is mediated by phospholipase C{gamma} and IP3R
Am J Physiol Cell Physiol,
December 1, 2004;
287(6):
C1667 - C1678.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. S. Argetsinger, J.-L. K. Kouadio, H. Steen, A. Stensballe, O. N. Jensen, and C. Carter-Su
Autophosphorylation of JAK2 on Tyrosines 221 and 570 Regulates Its Activity
Mol. Cell. Biol.,
June 1, 2004;
24(11):
4955 - 4967.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X. Chu, Q. Tong, J. Y. Cheung, J. Wozney, K. Conrad, V. Mazack, W. Zhang, R. Stahl, D. L. Barber, and B. A. Miller
Interaction of TRPC2 and TRPC6 in Erythropoietin Modulation of Calcium Influx
J. Biol. Chem.,
March 12, 2004;
279(11):
10514 - 10522.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Li, C. Miller, S. Hegde, and D. Wojchowski
Roles for an Epo Receptor Tyr-343 Stat5 Pathway in Proliferative Co-signaling with Kit
J. Biol. Chem.,
October 17, 2003;
278(42):
40702 - 40709.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X. Chu, J. Y. Cheung, D. L. Barber, L. Birnbaumer, L. I. Rothblum, K. Conrad, V. Abrasonis, Y.-m. Chan, R. Stahl, D. J. Carey, et al.
Erythropoietin Modulates Calcium Influx through TRPC2
J. Biol. Chem.,
September 6, 2002;
277(37):
34375 - 34382.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Haq, A. Halupa, B. K. Beattie, J. M. Mason, B. W. Zanke, and D. L. Barber
Regulation of Erythropoietin-induced STAT Serine Phosphorylation by Distinct Mitogen-activated Protein Kinases
J. Biol. Chem.,
May 3, 2002;
277(19):
17359 - 17366.
[Abstract]
[Full Text]
[PDF]
|
|
|
| |
