Identification of protein tyrosine kinases with oncogenic potential using a retroviral insertion mutagenesis screen.
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.
Department of Molecular and Developmental Genetics, VIB, Leuven, Belgium.
Els
E
Lierman
Helen
H
Van Miegroet
Els
E
Beullens
Jan
J
Cools
D002460:Cell Line; D002472:Cell Transformation, Viral / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D016254:Mutagenesis, Insertional / Q000379:methods*; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D012190:Retroviridae / Q000201:enzymology / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods
D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; D011505:Protein-Tyrosine Kinases