A growth signal with an artificially induced erythropoietin receptor-gp130 cytoplasmic domain heterodimer.
We report a strategy for generating efficient signal transduction with unnatural heterologous receptor combinations. As previously described [Ueda, H., Kawahara, M. et al. (2000) J. Immunol. Methods 241, 159-170], chimeric receptors composed of the V(H)/V(L) domains of anti-hen egg lysozyme antibody HyHEL-10 and N-terminally truncated erythropoietin receptor (EpoR) can be activated by lysozyme. When the cytoplasmic domains of these receptors were substituted with one derived from gp130, IL-3 dependent Ba/F3 cells expressing both V(H)-gp130 and V(L)-gp130 grew dose-dependently when given lysozyme without IL-3. However, cells expressing the heterologous pair of V(H)-gp130 and V(L)-EpoR also showed more efficient and stricter lysozyme-dependent proliferation in the absence of IL-3, indicating this combination is as an efficient and strict signal transducer as wild-type EpoR. The immunoprecipitation data indicated the existence of a preformed V(H)-gp130 and V(L)-EpoR heterodimer in the absence of lysozyme, suggesting the crucial role of a receptor conformational change in signal triggering as well as wild-type EpoR and gp130. Phosphorylation of JAK2, STAT3, and STAT5 was observed upon the addition of lysozyme, suggesting the activation of both EpoR- and gp130-derived signals.
Kawahara
M
M
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Ueda
H
H
Tsumoto
K
K
Kumagai
I
I
Mahoney
W
W
Nagamune
T
T
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D050822:Cytokine Receptor gp130; D019281:Dimerization; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D009113:Muramidase / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D017467:Receptors, Erythropoietin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*
D000906:Antibodies; D015703:Antigens, CD; C494200:Il6st protein, mouse; D008562:Membrane Glycoproteins; D017467:Receptors, Erythropoietin; D011993:Recombinant Fusion Proteins; D050822:Cytokine Receptor gp130; C421864:hen egg lysozyme; D009113:Muramidase