title: Growth hormone (GH)-independent dimerization of GH receptor by a leucine zipper results in constitutive activation.
abstract: Growth hormone initiates signaling by inducing homodimerization of two GH receptors. Here, we have sought to determine whether constitutively active receptor can be created in the absence of the extracellular domain by substituting it with high affinity leucine zippers to create dimers of the growth hormone receptor (GHR) signaling domain. The entire extracellular domain of the GHR was replaced by the hemagglutinin-tagged zipper sequence of either the c-Fos or c-Jun transcription factor (termed Fos-GHR and Jun-GHR, respectively). Transient transfection of Fos-GHR or Jun-GHR resulted in activation of the serine protease inhibitor 2.1 promoter in Chinese hamster ovary-K1 cells to a level equal to that achieved by fully activated wild type GHR. Furthermore, stable expression of Jun-GHR alone or Fos-GHR and Jun-GHR together in the interleukin 3-dependent BaF-B03 cell line resulted in cell proliferation after interleukin 3 withdrawal at a rate equal to maximally stimulated wild type GHR-expressing cells. Activation of STAT 5b was also observed in Fos-Jun-GHR-expressing cells at a level equal to that in chronically GH-treated GHR-expressing cells. Thus, forced dimerization of the transmembrane and cytoplasmic domains of the GHR in the absence of the extracellular domain can lead to the constitutive activation of known GH signaling end points, supporting the view that proximity of Janus kinase 2 (JAK2) kinases is the essential element in signaling. Such constitutively active GH receptors may have particular utility for transgenic livestock applications.
authors.affiliation: Physiology & Pharmacology Department and Centre for Molecular & Cellular Biology, University of Queensland, St. Lucia 4072, Australia.
authors.forename: S N
authors.initials: SN
authors.lastname: Behncken
authors.forename: N
authors.initials: N
authors.lastname: Billestrup
authors.forename: R
authors.initials: R
authors.lastname: Brown
authors.forename: J
authors.initials: J
authors.lastname: Amstrup
authors.forename: B
authors.initials: B
authors.lastname: Conway-Campbell
authors.forename: M J
authors.initials: MJ
authors.lastname: Waters
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002455:Cell Division; D006224:Cricetinae; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D019281:Dimerization; D013006:Growth Hormone / Q000378:metabolism*; D006801:Humans; D016350:Leucine Zippers; D008894:Milk Proteins; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D011986:Receptors, Somatotropin / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D050799:STAT5 Transcription Factor; D015534:Trans-Activators / Q000096:biosynthesis
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D008894:Milk Proteins; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D011986:Receptors, Somatotropin; D011993:Recombinant Fusion Proteins; D050799:STAT5 Transcription Factor; D015534:Trans-Activators; D013006:Growth Hormone