RET oncoproteins induce tyrosine phosphorylation changes of proteins involved in RNA metabolism

L. Gorla, M. Cantù, F. Miccichè, C. Patelli, P. Mondellini, M. A. Pierotti, I. Bongarzone

Research output: Contribution to journalArticle

Abstract

We report the identification of proteins induced in response to RET/PTC2, an oncogene implicated in thyroid cancers. Anti-phosphotyrosine antibody affinity resin was used to purify Tyr(P)-containing and interacting proteins from 293T and NIH3T3 cells which were transfected with kinase active or inactive RET/PTC and RETMEN2 oncogenes. Proteins were separated by one-dimensional SDS-PAGE, extracted by in-gel digestion, and identified by MALDI-TOF peptide mass fingerprinting. The expression and tyrosine phosphorylation of Sam68, a protein implicated in mRNA nucleocytoplasmic translocation and splicing, were further examined in RET-transfected cells and thyroid tumors. Of relevance, cells transfected with RETMEN2B examined for anti-phosphotyrosine bound proteins, showed other proteins implicated in splicing: DEAD-box p68 RNA helicase, SYNCRIP, and hnRNP K. Western blotting analysis suggested that these proteins are singularly tyrosine phosphorylated in RETMEN2B-transfected cells, and that they constitutively bind with Sam68. The study concludes that regulation of splicing factors is likely to be important in RET-mediated thyroid carcinogenesis.

Original languageEnglish
Pages (from-to)2272-2282
Number of pages11
JournalCellular Signalling
Volume18
Issue number12
DOIs
Publication statusPublished - Dec 2006

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Keywords

  • Oncogenic signalling network
  • RET
  • RET/PTC
  • Sam68
  • Thyroid tumors
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Cell Biology

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