Differential activation of Stat3 and Stat5 by distinct regions of the growth hormone receptor

The GH receptor (GHR) is a member of the cytokine receptor superfamily; its signaling involves the activation of Janus tyrosine kinases (JAK2) and Stat (signal transducers and activators of transcription) transcription factors. Using truncated and tyrosine mutants of the receptor, we show that different receptor domains are essential for the activation of Stat3 and Stat5. GH-dependent phosphorylation of JAK2, Stat3, and Stat5, as well as transactivation studies with reporter genes containing Stat3 and Stat5 DNA-binding elements, was performed in cells expressing the various GHR mutants. The membrane-proximal region of the receptor necessary for JAK2 activation is sufficient for Stat3 activation. In contrast, C-terminal tyrosine residues of GHR are absolutely required for Stat5 activation. The same residues are also involved in the regulation of JAK2 dephosphorylation, possibly through the activation of a phosphatase. Using in vitro experiments with glutathione-S-transferase fusion proteins, we demonstrate that the SH2 domain of Stat5 binds to the carboxy-terminal tyrosine-phosphorylated residues of GHR. Our results show that a cytokine receptor can mediate differently the activation of distinct Stat proteins that could be involved in cytokine-specific effects.