Identification of protein tyrosine kinases with oncogenic potential using a retroviral insertion mutagenesis screen

Haematologica. 2009 Oct;94(10):1440-4. doi: 10.3324/haematol.2009.007328.

Abstract

Protein tyrosine kinases form a large family of signaling proteins implicated in both normal and malignant cell signaling. The aim of this study was to identify protein tyro-sine kinases that can transform hematopoietic cells to growth factor independent proliferation when constitutively activated by homodimerization. We used a modified retroviral insertion mutagenesis screen with a retroviral vector containing the homodimerization domain of ETV6 followed by an artificial splice donor site. Integration of this retroviral vector within a gene of the host genome would generate a fusion transcript containing the dimerization domain and part of the disrupted gene. Using this strategy with the IL3 dependent Ba/F3 cell line, we identified 8 different protein tyrosine kinases (Abl1, Fgfr1, Hck, Jak2, Lck, Mertk, Mst1r, Tnk1) that transformed the cells. These results characterize HCK, MERTK, MST1R and TNK1 as potential oncogenes and describe a method to identify gain-of-function fusion genes using a retroviral insertion screen.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Transformation, Viral / genetics
  • Genetic Testing / methods
  • Humans
  • Mutagenesis, Insertional / methods*
  • Protein-Tyrosine Kinases / genetics*
  • Proto-Oncogene Proteins / genetics*
  • Recombinant Fusion Proteins / genetics*
  • Retroviridae / enzymology
  • Retroviridae / genetics*
  • Transduction, Genetic / methods

Substances

  • Proto-Oncogene Proteins
  • Recombinant Fusion Proteins
  • Protein-Tyrosine Kinases