In vivo targeting of the growth hormone receptor (GHR) Box1 sequence demonstrates that the GHR does not signal exclusively through JAK2.
GH is generally believed to signal exclusively through Janus tyrosine kinases (JAK), particularly JAK2, leading to activation of signal transducers and activators of transcription (STAT), ERK and phosphatidylinositol 3-kinase pathways, resulting in transcriptional regulation of target genes. Here we report the creation of targeted knock-in mice wherein the Box1 motif required for JAK2 activation by the GH receptor (GHR) has been disabled by four Pro/Ala mutations. These mice are unable to activate hepatic JAK2, STAT3, STAT5, or Akt in response to GH injection but can activate Src and ERK1/2. Their phenotype is identical to that of the GHR(-/-) mouse, emphasizing the key role of JAK2 in postnatal growth and the minimization of obesity in older males. In particular, they show dysregulation of the IGF-I/IGF-binding protein axis at transcript and protein levels and decreased bone length. Because no gross phenotypic differences were evident between GHR(-/-) and Box1 mutants, we undertook transcript profiling in liver from 4-month-old males. We compared their transcript profiles with our 391-GHR truncated mice, which activate JAK2, ERK1/2, and STAT3 in response to GH but not STAT5a/b. This has allowed us for the first time to identify in vivo Src/ERK-regulated transcripts, JAK2-regulated transcripts, and those regulated by the distal part of the GHR, particularly by STAT5.
Barclay
Johanna L
JL
Institute for Molecular Bioscience, University of Queensland, St. Lucia 4072, Australia. j.barclay@uq.edu.au
Kerr
Linda M
LM
Arthur
Leela
L
Rowland
Jennifer E
JE
Nelson
Caroline N
CN
Ishikawa
Mayumi
M
d'Aniello
Elisabetta M
EM
White
Mary
M
Noakes
Peter G
PG
Waters
Michael J
MJ
D000375:Aging; D000818:Animals; D001837:Body Weights and Measures; D002460:Cell Line; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D055879:Gene Knock-In Techniques; D013006:Growth Hormone / Q000494:pharmacology; D018969:Insulin-Like Growth Factor Binding Proteins / Q000235:genetics / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism; D053614:Janus Kinase 2 / Q000378:metabolism*; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009928:Organ Specificity; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011986:Receptors, Somatotropin / Q000235:genetics* / Q000378:metabolism*; D050791:STAT Transcription Factors / Q000378:metabolism; D012727:Sex Characteristics; D015398:Signal Transduction; D019061:src-Family Kinases / Q000378:metabolism
D018969:Insulin-Like Growth Factor Binding Proteins; D011986:Receptors, Somatotropin; D050791:STAT Transcription Factors; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2; D019061:src-Family Kinases; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases