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Cited by 10Receptor Mechanisms and SignalingPlatelet Disorders and TreatmentsProtein Structure and Dynamics

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Constitutive activation and oncogenicity are mediated by loss of helical structure at the cytosolic boundary of thrombopoietin receptor mutant dimers

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Defour, Jean-Philippe, Leroy, Emilie, Dass, Sharmila, Balligand, Thomas, Levy, Gabriel, Brett, Ian C, Papadopoulos, Nicolas, Mouton, Céline, Genet, Lidvine, Pecquet, Christian, Staerk, Judith, Smith, Steven O, Constantinescu, Stefan N·2023·eLife
DOI 10.7554/elife.81521PMC PMC10313314

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Abstract

Dimerization of the thrombopoietin receptor (TpoR) is necessary for receptor activation and downstream signaling through activated Janus kinase 2. We have shown previously that different orientations of the transmembrane (TM) helices within a receptor dimer can lead to different signaling outputs. Here we addressed the structural basis of activation for receptor mutations S505N and W515K that induce myeloproliferative neoplasms. We show using in vivo bone marrow reconstitution experiments that ligand-independent activation of TpoR by TM asparagine (Asn) substitutions is proportional to the proximity of the Asn mutation to the intracellular membrane surface. Solid-state NMR experiments on TM peptides indicate a progressive loss of helical structure in the juxtamembrane (JM) R/KWQFP motif with proximity of Asn substitutions to the cytosolic boundary. Mutational studies in the TpoR cytosolic JM region show that loss of the helical structure in the JM motif by itself can induce activation, but only when localized to a maximum of six amino acids downstream of W515, the helicity of the remaining region until Box 1 being required for receptor function. The constitutive activation of TpoR mutants S505N and W515K can be inhibited by rotation of TM helices within the TpoR dimer, which also restores helicity around W515. Together, these data allow us to develop a general model for activation of TpoR and explain the critical role of the JM W515 residue in the regulation of the activity of the receptor.

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Metadata

Authors
Defour, J.-P., Leroy, E., Dass, S., Balligand, T., Levy, G., Brett, I. C., Papadopoulos, N., Mouton, C., Genet, L., Pecquet, C., Staerk, J., Smith, S. O., & Constantinescu, S. N.
Affiliations
  • Ludwig Cancer Research (BE)
  • Stony Brook University (US)
Corresponding author
  • Smith, Steven O
  • Constantinescu, Stefan N
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2023-06-20
Volume
12
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.81521
PMC
PMC10313314
OpenAlex
W4381308829
Cited by
10
References
1
Citations by year
2026: 2 · 2025: 4 · 2024: 2 · 2023: 2
FWCI
0.89
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Access to Libraries (Université catholique de Louvain (UCL), l'Université de Namur (UNamur) and the Université Saint-Louis (USL-B)) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Fonds National de la Recherche Scientifique (Télévie PhD fellowship)
  • Les avions de Sebastien
  • Ludwig Institute for Cancer Research
  • Stichting Tegen Kanker
  • Catholic University of Louvain (Salus Sanguinis)
  • Les avions de Sébastien
  • Action de recherche concertée (16/21-073)
  • Walloon Excellence in Lifesciences and Biotechnology (F 44/8/5 - MCF/UIG - 10955)
  • W. M. Keck Foundation
  • Stony Brook University (National Institutes of Health, RO1 GM 46732)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Health Sciences › Medicine › Hematology › Platelet Disorders and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
thrombopoietin receptor, constitutive receptor activation, myeloproliferative neoplasms, juxtamembrane region, receptor dimerization, ligand-independent receptor activation, bone marrow reconstitution
MeSH terms
Cell Line, Mutation, Signal Transduction, Protein Structure, Secondary, Receptors, Thrombopoietin

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