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Analysis of transcription and protein changes in response to human RET activation in SKN‐SH and transfected COS cells.
Author(s) -
Torres Martha Angelina,
Poku Maryann,
Vega Quinn
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.829.2
Subject(s) - internalization , proto oncogene proteins c ret , gene isoform , biology , receptor tyrosine kinase , tyrosine kinase , microbiology and biotechnology , transcription factor , glial cell line derived neurotrophic factor , transfection , receptor , kinase , signal transduction , cancer research , cell culture , gene , genetics , neurotrophic factors
RET is a mammalian receptor tyrosine kinase normally activated after association with a protein co‐receptor GFR‐a (1–4) and its corresponding ligand (GDNF, Neurturin, Artemin or Persephin). RET is required for nervous system development and RET mutations are associated with both cancer (MEN2A, MEN2B and FMTC) and developmental abnormalities (Hirschsprung's Disease). In order to analyze how the cell responds to RET activation, isoform expression, internalization and downstream transcription were measured in RET expressing cells. QPCR was used to measure the production of the two major isoforms of RET‐RET9 and RET51. In these experiments, RET 9 levels increased ∼ 4‐fold in comparison to RET 51. With respect to internalization, the loss of RET was identified over time with a loss of the RET protein by 6 hours. With respect to signaling, downstream luciferase promoter assays demonstrated that wild type RET increased transcription over phosphotyrosine mutants and kinase inactive RET. Future experiments will confirm these changes and study the effect of both specific tyrosines and naturally occurring mutations on RET signaling. (Supported by NIH Grant R15‐NS048043)