Mechanism of activation of protein kinase JAK2 by the growth hormone receptor.
Signaling from JAK (Janus kinase) protein kinases to STAT (signal transducers and activators of transcription) transcription factors is key to many aspects of biology and medicine, yet the mechanism by which cytokine receptors initiate signaling is enigmatic. We present a complete mechanistic model for activation of receptor-bound JAK2, based on an archetypal cytokine receptor, the growth hormone receptor. For this, we used fluorescence resonance energy transfer to monitor positioning of the JAK2 binding motif in the receptor dimer, substitution of the receptor extracellular domains with Jun zippers to control the position of its transmembrane (TM) helices, atomistic modeling of TM helix movements, and docking of the crystal structures of the JAK2 kinase and its inhibitory pseudokinase domain with an opposing kinase-pseudokinase domain pair. Activation of the receptor dimer induced a separation of its JAK2 binding motifs, driven by a ligand-induced transition from a parallel TM helix pair to a left-handed crossover arrangement. This separation leads to removal of the pseudokinase domain from the kinase domain of the partner JAK2 and pairing of the two kinase domains, facilitating trans-activation. This model may well generalize to other class I cytokine receptors.
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia. m.waters@uq.edu.au a.brooks@uq.edu.au.
Andrew J
AJ
Brooks
Department of Chemical and Biomolecular Engineering, University of Houston, Houston, TX 77004, USA.
Wei
W
Dai
The University of Queensland, School of Chemistry and Molecular Biosciences, St Lucia, Queensland 4072, Australia.
Megan L
ML
O'Mara
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Daniel
D
Abankwa
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Yash
Y
Chhabra
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Rebecca A
RA
Pelekanos
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Olivier
O
Gardon
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Kathryn A
KA
Tunny
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Kristopher M
KM
Blucher
Biota Structural Biology Laboratory and Australian Cancer Research Foundation (ACRF) Rational Drug Discovery Centre, St Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia.
Craig J
CJ
Morton
Biota Structural Biology Laboratory and Australian Cancer Research Foundation (ACRF) Rational Drug Discovery Centre, St Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia. Department of Biochemistry and Molecular Biology and Bio21 Institute, University of Melbourne, Parkville, Victoria 3052, Australia.
Michael W
MW
Parker
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Emma
E
Sierecki
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Yann
Y
Gambin
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Guillermo A
GA
Gomez
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia.
Kirill
K
Alexandrov
Scripps Research Institute, La Jolla, CA 92037, USA.
Ian A
IA
Wilson
Department of Chemical and Biomolecular Engineering, University of Houston, Houston, TX 77004, USA.
Manolis
M
Doxastakis
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia. The University of Queensland, School of Chemistry and Molecular Biosciences, St Lucia, Queensland 4072, Australia.
Alan E
AE
Mark
The University of Queensland, Institute for Molecular Bioscience (IMB), St Lucia, Queensland 4072, Australia. m.waters@uq.edu.au a.brooks@uq.edu.au.
Michael J
MJ
Waters
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D003545:Cysteine / Q000737:chemistry; D004789:Enzyme Activation; D057809:HEK293 Cells; D006801:Humans; D053614:Janus Kinase 2 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011986:Receptors, Somatotropin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D011986:Receptors, Somatotropin; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D003545:Cysteine