J. Fujimoto et al., CHARACTERIZATION OF THE TRANSFORMING ACTIVITY OF P80, A HYPERPHOSPHORYLATED PROTEIN IN A KI-1 LYMPHOMA CELL-LINE WITH CHROMOSOMAL TRANSLOCATION T(2-5), Proceedings of the National Academy of Sciences of the United Statesof America, 93(9), 1996, pp. 4181-4186
We have molecularly cloned a cDNA encoding a protein uniquely expresse
d and hyperphosphorylated at tyrosine residues in a Ki-1 lymphoma cell
that contained chromosomal translocation t(2;5). The encoded protein
p80 was shown to be generated by fusion of a protein-tyrosine kinase a
nd a nucleolar protein B23/nucleophosmin (NPM). The coding sequence of
this cDNA turned out to be virtually identical to that of the fusion
cDNA for NPM-anaplastic lymphoma kinase (ALK) previously cloned from t
he transcript of the gene at the breakpoint of the same translocation.
Overexpression of p80 in NM 3T3 cells induced neoplastic transformati
on, suggesting that the p80 kinase is aberrantly activated. The normal
form of p80 was predicted to be a receptor type tyrosine kinase on th
e basis of its sequence similarity to the insulin receptor family of k
inases. However, an immunofluorescence study using COS cells revealed
that p80 was localized to the cytoplasm. Thus, subcellular translocati
on and activation of the tyrosine kinase presumably by its structural
alteration would cause the malignant transformation. We also showed th
at a mutant p80 lacking the NPM portion was unable to transform NM 3T3
cells. Thus, the NPM sequence is essential for the transforming activ
ity, suggesting that the chromosomal translocation is responsible for
the oncogenesis. Finally, She and insulin receptor substrate 1 (IRS-1)
were tyrosine-phosphorylated and bound to p80 in p80-transformed cell
s. However, mutants of p80 that were defective for binding to and phos
phorylation of She and insulin receptor substrate 1 could transform NM
3T3 cells. Association of these mutants with GRB2 was still observed,
suggesting that interaction of p80 with GRB2 but not with She or IRS-
1 was relevant for cell transformation.