Regulation of signal transducer and activator of transcription (STAT) 5b activation by the temporal pattern of growth hormone stimulation.

Plasma GH profiles, intermittent in adult male and continuous in adult female rats, respectively, activate unique patterns of gene transcription in male and female rat liver. Pulsatile, but not continuous, GH exposure activates liver STAT5 (signal transducer and activator of transcription-5) by tyrosine phosphorylation, leading to nuclear translocation, and is proposed to play a key role in GH pulse-regulated male-specific liver gene expression. The mechanisms underlying the GH pattern dependence of STAT5 activation are presently investigated using a rat hepatocyte-derived cell line. Rat GH stimulated tyrosine phosphorylation followed by serine or threonine phosphorylation, leading to activation of the DNA-binding activity of STAT5b, the major STAT5 form present in these cells. Maximal STAT5b activation required a full 20 min at a receptor-saturating GH concentration of 50 ng/ml, suggesting that hormone binding leading to receptor dimerization is a relatively slow process. Repeat cycles of GH pulsation led to repeat cycles of STAT5b activation followed by deactivation, similar to rat liver in vivo. Full responsiveness to succeeding GH pulses required a minimum GH off-time of > or = 2.5 h, but was independent of new protein synthesis. Continuous GH exposure led to down-regulation of activated STAT5b, consistent with the desensitization of this GH pulse-activated pathway observed in female rat liver. The rapid deactivation of STAT5b after termination of a GH pulse involved phosphotyrosine dephosphorylation as a key first step and could be blocked by pervanadate, a phosphotyrosine phosphatase inhibitor. Unexpectedly, serine/threonine kinase inhibitors also inhibited STAT5b deactivation. These studies establish that STAT5b is responsive to the temporal pattern of GH stimulation and demonstrate a role for both a tyrosine phosphatase and a serine/threonine kinase in resetting this JAK/STAT signaling apparatus so that it may respond to subsequent rounds of GH pulse activation.