Jackson P K, Paskind M, Baltimore D
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142.
Oncogene. 1993 Jul;8(7):1943-56.
c-abl is the normal cellular homolog of the v-abl transforming gene of Abelson murine leukemia virus. By constructing recombinants between c- and v-abl retroviruses, we show that a point mutation in c-Abl is sufficient to change the myristoylated form of c-Abl into a protein able to transform fibroblasts, but not capable of transforming bone marrow or inducing Abelson disease. This activating mutation, which changes the phenylalanine at amino acid 420 to valine (F420V) found in the homologous position of v-Abl, is positioned outside of the SH3 domain, a region typically modified in transforming alleles of abl. Phenylalanine 420 is perfectly conserved among tyrosine kinases with N-terminal SH3 domains (the Src and Abl families). The equivalent position in other protein tyrosine kinases is a conserved hydrophobic residue that predicts the specific family to which that kinase belongs. Mutation of phenylalanine 420 to other hydrophobic residues activates c-Abl. Unlike other transforming variants of Abl, the F420V mutant protein is not highly phosphorylated on tyrosine. Mutation of the nearby proposed autophosphorylation site, tyrosine 412, shows that this tyrosine is not strictly required for fibroblast transformation in either F420V or SH3-deleted variants of c-Abl (IV).
c-abl是阿贝尔森鼠白血病病毒v-abl转化基因的正常细胞同源物。通过构建c-abl和v-abl逆转录病毒之间的重组体,我们发现c-Abl中的一个点突变足以将肉豆蔻酰化形式的c-Abl转变为一种能够转化成纤维细胞的蛋白质,但不能转化骨髓或引发阿贝尔森病。这种激活突变将第420位氨基酸的苯丙氨酸变为缬氨酸(F420V),该突变位于v-Abl同源位置,位于SH3结构域之外,SH3结构域是abl转化等位基因中通常被修饰的区域。苯丙氨酸420在具有N端SH3结构域的酪氨酸激酶(Src和Abl家族)中完全保守。其他蛋白酪氨酸激酶中的等效位置是一个保守的疏水残基,可预测该激酶所属的特定家族。将苯丙氨酸420突变为其他疏水残基可激活c-Abl。与Abl的其他转化变体不同,F420V突变蛋白在酪氨酸上的磷酸化程度不高。对附近假定的自磷酸化位点酪氨酸412进行突变表明,在c-Abl的F420V或SH3缺失变体(IV)中,该酪氨酸对于成纤维细胞转化并非严格必需。