Direct phosphorylation of proliferative and survival pathway proteins by RET

Ganesh R. Panta, Lihua Du, Fiemu E. Nwariaku, Lawrence T. Kim

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Background. Gain-of-function mutations in the RET tyrosine kinase receptor cause the multiple endocrine neoplasia syndromes type 2a and 2b, and medullary thyroid cancer. We have previously shown that RET signals through focal adhesion kinase (FAK) in medullary thyroid cancer cells and that extracellular signal-regulated kinase (ERK) activity can be blocked by pp2, an inhibitor of both Src and RET. We hypothesized that RET could directly phosphorylate FAK and ERK. Methods. RET and ERK kinase activity were measured with the use of an in vitro kinase assay. The relative contribution of RET in phosphorylation of ERK was tested by treating cells with PD98059, an inhibitor of MEK, and the RET inhibitor PP2, then measuring ERK activity. Results. Immunoprecipitated, mutant RET from cells or the recombinant RET kinase domain was able to directly phosphorylate tyrosine residues on FAK. Specifically Y576/577, Y861, and Y925, but not the autophosphorylation site Y397 of FAK, were phosphorylated by RET. Similarly ERK 2 could be phosphorylated at Y187 (Y204 in ERK1). Inhibition of both MEK (upstream of ERK) and RET was more potent than inhibition of either alone in decreasing ERK activity. Furthermore, tyrosine residues in DOK1, the p85 subunit of phosphatidylinositol 3′ kinase, JNK 1 and 2, P-38, and phospholipase-γ were directly phosphorylated by RET. Conclusions. RET directly phosphorylates tyrosine residues on FAK, ERK 1/2, DOK1, the p85 subunit of of phosphatidylinositol 3′ kinase, JNK 1 and 2, P-38, and phospholipase-γ. These data indicate a direct interaction between RET and a broad range of effector molecules that may contribute to tumor pathogenesis.

Original languageEnglish (US)
Pages (from-to)269-274
Number of pages6
JournalSurgery
Volume138
Issue number2
DOIs
StatePublished - Aug 1 2005

ASJC Scopus subject areas

  • Surgery

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