title: Centrosome-kinase fusions promote oncogenic signaling and disrupt centrosome function in myeloproliferative neoplasms.
abstract: Chromosomal translocations observed in myeloproliferative neoplasms (MPNs) frequently fuse genes that encode centrosome proteins and tyrosine kinases. This causes constitutive activation of the kinase resulting in aberrant, proliferative signaling. The function of centrosome proteins in these fusions is not well understood. Among others, kinase centrosome localization and constitutive kinase dimerization are possible consequences of centrosome protein-kinase fusions. To test the relative contributions of localization and dimerization on kinase signaling, we targeted inducibly dimerizable FGFR1 to the centrosome and other subcellular locations and generated a mutant of the FOP-FGFR1 MPN fusion defective in centrosome localization. Expression in mammalian cells followed by western blot analysis revealed a significant decrease in kinase signaling upon loss of FOP-FGFR1 centrosome localization. Kinase dimerization alone resulted in phosphorylation of the FGFR1 signaling target PLCγ, however levels comparable to FOP-FGFR1 required subcellular targeting in addition to kinase dimerization. Expression of MPN fusion proteins also resulted in centrosome disruption in epithelial cells and transformed patient cells. Primary human MPN cells showed masses of modified tubulin that colocalized with centrin, Smoothened (Smo), IFT88, and Arl13b. This is distinct from acute myeloid leukemia (AML) cells, which are not associated with centrosome-kinase fusions and had normal centrosomes. Our results suggest that effective proliferative MPN signaling requires both subcellular localization and dimerization of MPN kinases, both of which may be provided by centrosome protein fusion partners. Furthermore, centrosome disruption may contribute to the MPN transformation phenotype.
authors.affiliation: Department of Biology, Stanford University, Stanford, California, United States of America.
authors.forename: Joanna Y
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Stanford Institute for Stem Cell Biology and Regenerative Medicine and Cancer Institute, Stanford University School of Medicine, Stanford, California, United States of America; Department of Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Wan-Jen
authors.initials: WJ
authors.lastname: Hong
authors.affiliation: Stanford Institute for Stem Cell Biology and Regenerative Medicine and Cancer Institute, Stanford University School of Medicine, Stanford, California, United States of America; Department of Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Ravindra
authors.initials: R
authors.lastname: Majeti
authors.affiliation: Department of Biology, Stanford University, Stanford, California, United States of America; Department of Genetics, Stanford School of Medicine, Stanford, California, United States of America.
authors.forename: Tim
authors.initials: T
authors.lastname: Stearns
mesh_terms: D018385:Centrosome / Q000378:metabolism*; D015870:Gene Expression; D023061:Gene Order; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000235:genetics* / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics; D015514:Oncogene Proteins, Fusion / Q000235:genetics* / Q000378:metabolism*; D051966:Phospholipase C gamma / Q000378:metabolism; D010766:Phosphorylation; D055503:Protein Multimerization; D021381:Protein Transport; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics
chemical_list: C541075:CHTOP protein, human; D009687:Nuclear Proteins; D015514:Oncogene Proteins, Fusion; D014157:Transcription Factors; D011505:Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D051966:Phospholipase C gamma