TLS/FUS, a pro-oncogene involved in multiple chromosomal translocations, is a novel regulator of BCR/ABL-mediated leukemogenesis.
The leukemogenic potential of BCR/ABL oncoproteins depends on their tyrosine kinase activity and involves the activation of several downstream effectors, some of which are essential for cell transformation. Using electrophoretic mobility shift assays and Southwestern blot analyses with a double-stranded oligonucleotide containing a zinc finger consensus sequence, we identified a 68 kDa DNA-binding protein specifically induced by BCR/ABL. The peptide sequence of the affinity-purified protein was identical to that of the RNA-binding protein FUS (also called TLS). Binding activity of FUS required a functional BCR/ABL tyrosine kinase necessary to induce PKCbetaII-dependent FUS phosphorylation. Moreover, suppression of PKCbetaII activity in BCR/ABL-expressing cells by treatment with the PKCbetaII inhibitor CGP53353, or by expression of a dominant-negative PKCbetaII, markedly impaired the ability of FUS to bind DNA. Suppression of FUS expression in myeloid precursor 32Dcl3 cells transfected with a FUS antisense construct was associated with upregulation of the granulocyte-colony stimulating factor receptor (G-CSFR) and downregulation of interleukin-3 receptor (IL-3R) beta-chain expression, and accelerated G-CSF-stimulated differentiation. Downregulation of FUS expression in BCR/ABL-expressing 32Dcl3 cells was associated with suppression of growth factor-independent colony formation, restoration of G-CSF-induced granulocytic differentiation and reduced tumorigenic potential in vivo. Together, these results suggest that FUS might function as a regulator of BCR/ABL leukemogenesis, promoting growth factor independence and preventing differentiation via modulation of cytokine receptor expression.
Perrotti
D
D
Department of Microbiology and Immunology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA. perrott1@jeflin.tjn.edu
Bonatti
S
S
Trotta
R
R
Martinez
R
R
Skorski
T
T
Salomoni
P
P
Grassilli
E
E
Lozzo
R V
RV
Cooper
D R
DR
Calabretta
B
B
D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D016044:Fusion Proteins, bcr-abl / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic; D006133:Growth Substances / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D007951:Leukemia, Myeloid / Q000201:enzymology / Q000209:etiology / Q000235:genetics*; D051379:Mice; D008813:Mice, Inbred ICR; D016513:Mice, SCID; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011493:Protein Kinase C / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis; D011519:Proto-Oncogenes / Q000502:physiology*; D034702:RNA-Binding Protein FUS; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D014178:Translocation, Genetic; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D006133:Growth Substances; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D034702:RNA-Binding Protein FUS; D012261:Ribonucleoproteins; D011505:Protein-Tyrosine Kinases; D016044:Fusion Proteins, bcr-abl; D011493:Protein Kinase C