Department of Molecular and Developmental Genetics, VIB, Leuven, Belgium.
Haematologica. 2009 Oct;94(10):1440-4. doi: 10.3324/haematol.2009.007328.
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.
蛋白酪氨酸激酶形成了一个庞大的信号蛋白家族，涉及正常和恶性细胞信号。本研究的目的是鉴定蛋白酪氨酸激酶，当它们通过同源二聚化被持续激活时，能够将造血细胞转化为生长因子非依赖性增殖。我们使用了一种改良的逆转录病毒插入诱变筛选方法，使用包含 ETV6 同源二聚化结构域的逆转录病毒载体，随后是一个人工剪接供体位点。该逆转录病毒载体在宿主基因组内的基因中的整合将产生一个包含二聚化结构域和部分破坏基因的融合转录本。使用这种策略和 IL3 依赖性 Ba/F3 细胞系，我们鉴定出了 8 种不同的蛋白酪氨酸激酶（Abl1、Fgfr1、Hck、Jak2、Lck、Mertk、Mst1r、Tnk1），它们可以转化细胞。这些结果将 HCK、MERTK、MST1R 和 TNK1 表征为潜在的致癌基因，并描述了一种使用逆转录病毒插入筛选来鉴定获得功能融合基因的方法。
