McWhirter J R, Galasso D L, Wang J Y
Department of Biology, University of California, San Diego, La Jolla 92093-0116.
Mol Cell Biol. 1993 Dec;13(12):7587-95. doi: 10.1128/mcb.13.12.7587-7595.1993.
In Philadelphia chromosome-positive human leukemias, the c-abl proto-oncogene on chromosome 9 becomes fused to the bcr gene on chromosome 22, and chimeric Bcr-Abl proteins are produced. The fused Bcr sequences activate the tyrosine kinase, actin-binding, and transforming functions of Abl. Activation of the Abl transforming function has been shown to require two distinct domains of Bcr: domain 1 (Bcr amino acids 1 to 63) and domain 2 (Bcr amino acids 176 to 242). The amino acid sequence of domain 1 indicates that it may be a coiled-coil oligomerization domain. We show here that domain 1 of Bcr forms a homotetramer. Tetramerization of Bcr-Abl through Bcr domain 1 correlates with activation of the tyrosine kinase and F-actin-binding functions of Abl. Disruption of the coiled coil by insertional mutagenesis inactivates the oligomerization function as well as the ability of Bcr-Abl to transform Rat-1 fibroblasts or to abrogate interleukin-3 dependence in lymphoid cells. These results strongly suggest that Bcr-Abl oligomers are the active entities in transformation.
在费城染色体阳性的人类白血病中,9号染色体上的c-abl原癌基因与22号染色体上的bcr基因融合,产生嵌合的Bcr-Abl蛋白。融合的Bcr序列激活了Abl的酪氨酸激酶、肌动蛋白结合和转化功能。已证明激活Abl转化功能需要Bcr的两个不同结构域:结构域1(Bcr氨基酸1至63)和结构域2(Bcr氨基酸176至242)。结构域1的氨基酸序列表明它可能是一个卷曲螺旋寡聚化结构域。我们在此表明Bcr的结构域1形成同四聚体。通过Bcr结构域1实现的Bcr-Abl四聚化与Abl的酪氨酸激酶和F-肌动蛋白结合功能的激活相关。通过插入诱变破坏卷曲螺旋会使寡聚化功能以及Bcr-Abl转化大鼠-1成纤维细胞或消除淋巴细胞中白细胞介素-3依赖性的能力失活。这些结果强烈表明Bcr-Abl寡聚体是转化中的活性实体。