Daple-FLT3 (CCDC88C-FLT3) gene fusion requires the coiled-coil domain for maximal activation and pericentrosomal localization

Abstract

Gene fusions are stable protein products often occurring from chromosomal rearrangements. These chimeric proteins typically contain distinct molecular entities from each parent gene, and thus, create a product with altered or aberrant function. Gene fusions are frequently found in cancers, including Leukemia. Here, we characterize the kinase activity and subcellular distribution of the Daple-FLT3 (CCDC88C-FLT3) fusion oncoprotein--a rare, but recurrent gene fusion found in patients with hematological malignancies. The protein contains the FLT3 kinase domain and is activated without ligand stimulation. This leads to activation in STAT5a, AKT, and MAPK signaling, which can be modulated by the tyrosine kinase inhibitors (TKIs) sorafenib, quizartinib, and to a lesser degree, imatinib. Moreover, fusion of this kinase domain to Daple facilitates its localization to the pericentrosomal space and enhances kinase activation. These findings provide evidence that targeting Daple-FLT3 outside of its kinase domain may be a complementary approach with TKI therapy. Key PointsO_LIDaple-FLT3 fusion proteins contain a constitutively active kinase domain, activating distinct signaling molecules in cells C_LIO_LICoiled-coil domain on Daple is dispensable for kinase activation, but necessary for maximal activation C_LI