Role of TFG sequences outside the coiled-coil domain in TRK-T3 oncogenic activation.
The TRK-T3 oncoprotein, isolated from a human papillary thyroid tumor, arises from the fusion between the N-terminal domain of the TFG gene and the tyrosine kinase domain of the NTRK1 receptor. The 68 kDa TRK-T3 oncoprotein displays a constitutive tyrosine kinase activity resulting in its capability to transform NIH3T3 cells. The TFG portion of TRK-T3 contains a coiled-coil domain, which mediates protein oligomerization essential for the oncogene constitutive activation, and several consensus sites for protein interaction. In this study, we investigate the role of TFG sequences outside the coiled-coil domain on TRK-T3 activation, We constructed four mutants carrying different deletions of TFG sequences and expressed them in mammalian cells. By performing biochemical and biological assays we demonstrated that all the deleted regions are required for TRK-T3 activation, as they are involved in different mechanisms such as protein processing, formation of stable and/or functional complexes, and possible interaction with other proteins. By constructing site-specific mutants, we demonstrated a crucial role for a PB1 domain and a considerable contribution of an SH2-binding motif in TRK-T3 oncogenic activation. This work establishes an important role for TFG sequences outside the coiled-coil domain in the activation of the thyroid TRK-T3 oncogene.
Roccato
Emanuela
E
Operative Unit #3, Department of Experimental Oncology, Instituto Nazionale Tumori, Via G Venezian, Milan, Italy.
Pagliardini
Sonia
S
Cleris
Loredana
L
Canevari
Silvana
S
Formelli
Franca
F
Pierotti
Marco A
MA
Greco
Angela
A
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009374:Neoplasms, Experimental / Q000209:etiology / Q000235:genetics; D009857:Oncogenes / Q000502:physiology*; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D020917:Receptor, trkA / Q000235:genetics; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D033921:Vesicular Transport Proteins
D011506:Proteins; D020917:Receptor, trkA; C484335:Tfg protein, mouse; D033921:Vesicular Transport Proteins