Symmetric signaling by an asymmetric 1 erythropoietin: 2 erythropoietin receptor complex.
Via sites 1 and 2, erythropoietin binds asymmetrically to two identical receptor monomers, although it is unclear how asymmetry affects receptor activation and signaling. Here we report the design and validation of two mutant erythropoietin receptors that probe the role of individual members of the receptor dimer by selectively binding either site 1 or site 2 on erythropoietin. Ba/F3 cells expressing either mutant receptor do not respond to erythropoietin, but cells co-expressing both receptors respond to erythropoietin by proliferation and activation of the JAK2-Stat5 pathway. A truncated receptor with only one cytosolic tyrosine (Y343) is sufficient for signaling in response to erythropoietin, regardless of the monomer on which it is located. Similarly, only one receptor in the dimer needs a juxtamembrane hydrophobic L253 or W258 residue, essential for JAK2 activation. We conclude that despite asymmetry in the ligand-receptor interaction, both sides are competent for signaling, and appear to signal equally.
Zhang
Yingxin L
YL
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Radhakrishnan
Mala L
ML
Lu
Xiaohui
X
Gross
Alec W
AW
Tidor
Bruce
B
Lodish
Harvey F
HF
D001665:Binding Sites; D049109:Cell Proliferation; D002478:Cells, Cultured; D003198:Computer Simulation; D004921:Erythropoietin / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D017467:Receptors, Erythropoietin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism
D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D050799:STAT5 Transcription Factor; D004921:Erythropoietin; D014443:Tyrosine; D053614:Janus Kinase 2