Functional heterodimerization of prolactin and growth hormone receptors byovine placental lactogen

Although homo- or heterodimerization are common mechanisms for activation o f cytokine receptors, crosstalk. between two distinct receptors in this sup erfamily has been never shown. Here we show a physiologically relevant exam ple indicating that such an interaction does occurs, thus raising the hypot hesis that heterodimerization between distinct cytokine receptors may be a novel mechanism contributing to the diversity of cytokine signaling. These findings were documented using both surface plasmon resonance and gel filtr ation experiments and show that ovine placental lactogen (PL) heterodimeriz es the extracellular domains (ECDs) of ruminant growth hormone receptor (GH R) and prolactin receptor (PRLR). We also show that PL or PL analogues that exhibit little or no activity in cells transfected with PRLRs and no activ ity in cells transfected with ovine GHRs exhibit largely enhanced activity in cells cotransfected with both PRLRs and GHRs. Furthermore, chimeric rece ptors consisting of cytosolic and transmembrane part of ovine GHR or ovine PRLR and ECDs of human granulocyte-macrophage colony-stimulating factor rec eptor (GM-CSFR) alpha or beta were constructed. Upon transfection into Chin ese hamster ovary cells along with reporter luciferase gene and stimulation by GM-CSF, a significant increase in luciferase activity occurred when GM- CCFR-alpha-PRLR and GM-CSFR-beta-GHR or GM-CSFR-alpha-GHR and GM-CSRR-beta- PRLR were cotransfected. In conclusion, we show that ovine PL is capable of functional heterodimerization of GHR and PRLR and that when their cytosoli c parts, coupled to the ECD of GM-CSF receptors, are heterodimerized by GM- CSF, they are capable of transducing biological signal.