RET-familial medullary thyroid carcinoma mutants Y791F and S891A activate a Src/JAK/STAT3 pathway, independent of glial cell line-derived neurotrophic factor

Ivan Plaza Menacho, R Koster, Almer M. van der Sloot, WJ Quax, Jan Osinga, Tineke van der Sluis, HHarry Hollema, Grzegorz M Burzynski, O Gimm, Charles HCM Buys, Bart JL Eggen, Robert MW Hofstra

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76 Citations (Scopus)

Abstract

The RET proto-oncogene encodes a receptor tyrosine kinase whose dysfunction plays a crucial role in the development of several neural crest disorders. Distinct activating RET mutations cause multiple endocrine neoplasia type 2A (MEN2A), type 2B (MEN2B), and familial medullary thyroid carcinoma (FMTC). Despite clear correlations between the mutations found in these cancer syndromes and their phenotypes, the molecular mechanisms connecting the mutated receptor to the different disease phenotypes are far from completely understood. Luciferase reporter assays in combination with immunoprecipitations, and Western and immunohistochemistry analyses, were done in order to characterize the signaling properties of two FMTC-associated RET mutations, Y791F and S891A, respectively, both affecting the tyrosine kinase domain of the receptor. We show that these RET-FMTC mutants are monomeric receptors which are autophosphorylated and activated independently of glial cell line-derived neurotrophic factor. Moreover, we show that the dysfunctional signaling properties of these mutants, when compared with wild-type RET, involve constitutive activation of signal transducer and activator of transcription 3 (STAT3). Furthermore, we show that STAT3 activation is mediated by a signaling pathway involving Src, JAK1, and JAK2, differing from STAT3 activation promoted by RETC634R which was previously found to be independent of Src and JAKs. Three-dimensional modeling of the RET catalytic domain suggested that the structural changes promoted by the respective amino acids substitutions lead to a more accessible substrate and ATP-binding monomeric conformation. Finally, immunohistochemical analysis of FMTC tumor samples support the in vitro data, because nuclear localized, Y705-phosphorylated STAT3 as well as a high degree of RET expression at the plasma membrane was observed.

Original languageEnglish
Pages (from-to)1729-1737
Number of pages9
JournalCancer Research
Volume65
Issue number5
DOIs
Publication statusPublished - 1-Mar-2005

Keywords

  • RECEPTOR TYROSINE KINASE
  • MICE LACKING GDNF
  • AUTOPHOSPHORYLATION SITES
  • HIRSCHSPRUNGS-DISEASE
  • CRYSTAL-STRUCTURE
  • INSULIN-RECEPTOR
  • PROTEIN
  • MUTATIONS
  • PROTOONCOGENE
  • DOMAIN

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