Induction of tyrosine phosphorylation by the erythropoietin receptor correlates with mitogenesis

Osamu Miura, Alan D'Andrea, David Kabat, James N. Ihle

Research output: Contribution to journalArticle

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Abstract

A role for tyrosine phosphorylation in the signal-transducing mechanisms of several hematopoietic growth factors has been hypothesized. To extend these observations, we have examined the effects of erythropoietin (Epo) on tyrosine phosphorylation in an Epo-responsive cell that was obtained by transfecting the murine erythropoietin receptor (EpoR) into an interleukin-3 (IL-3)-dependent cell line. By two-dimensional analysis of phosphotyrosine-containing proteins isolated with a monoclonal antibody (1G2) against phosphotyrosine, Epo and IL-3 were found to rapidly induce tyrosine phosphorylation of comparable substrates of 92, 70, and 56 kDa. In addition, Epo uniquely induced phosphorylation of a 72-kDa substrate while IL-3 uniquely induced phosphorylation of a 140-kDa substrate. Immunoprecipitation and mixing experiments indicated that the 72-kDa substrate may represent a small fraction of the EpoR. To explore the significance of tyrosine phosphorylation, we generated two mutants of the EpoR that lacked 108 or 146 amino acids at their carboxyl termini. In addition we constructed an internally deleted mutant that lacked 20 amino acids in a region of sequence homology with the IL-2 receptor β chain. Although all mutants were expressed at comparable levels and had comparable binding affinities for Epo, only the mutant lacking 108 amino acids at the carboxyl terminus retained significant mitogenic activity or the ability to induce tyrosine phosphorylation.

Original languageEnglish (US)
Pages (from-to)4895-4902
Number of pages8
JournalMolecular and Cellular Biology
Volume11
Issue number10
StatePublished - 1991
Externally publishedYes

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Erythropoietin Receptors
Tyrosine
Phosphorylation
Erythropoietin
Interleukin-3
Phosphotyrosine
Amino Acids
Interleukin-2 Receptors
Sequence Homology
Immunoprecipitation
Intercellular Signaling Peptides and Proteins
Monoclonal Antibodies
Cell Line

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Induction of tyrosine phosphorylation by the erythropoietin receptor correlates with mitogenesis. / Miura, Osamu; D'Andrea, Alan; Kabat, David; Ihle, James N.

In: Molecular and Cellular Biology, Vol. 11, No. 10, 1991, p. 4895-4902.

Research output: Contribution to journalArticle

Miura, O, D'Andrea, A, Kabat, D & Ihle, JN 1991, 'Induction of tyrosine phosphorylation by the erythropoietin receptor correlates with mitogenesis', Molecular and Cellular Biology, vol. 11, no. 10, pp. 4895-4902.
Miura, Osamu ; D'Andrea, Alan ; Kabat, David ; Ihle, James N. / Induction of tyrosine phosphorylation by the erythropoietin receptor correlates with mitogenesis. In: Molecular and Cellular Biology. 1991 ; Vol. 11, No. 10. pp. 4895-4902.
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