Amson R B, Marcelle C, Telerman A
Institute of Interdisciplinary Research, School of Medicine, Free University of Brussels, Belgium.
Oncogene. 1989 Feb;4(2):243-7.
Human chronic myelogenous leukemia is characterized by a reciprocal translocation between chromosomes 9 and 22. This results in the transfer of the c-abl protooncogene from chromosome 9 into the bcr gene on chromosome 22. The purpose of this study was to characterize the bcr and related gene products. Antibodies were raised against a fused trpE-bcr protein induced in a bacterial expression vector. Immunoprecipitation with the monoclonal and polyclonal antibodies of metabolically [35S]methionine labeled leukemic cell lines shows a 210, 160 and 130 Kda protein in Philadelphia positive cells containing the bcr-abl fused transcript. Only the 160 and 130 Kda were present in the Philadelphia negative cells. In vitro kinase assay shows that the 130 Kda protein is a phosphoprotein mainly phosphorylated on serine. Partial proteolysis indicates that the p210 and p130 share common domains. In subcellular fractionation experiments, the p130 is colocalized with the p210 bcr-abl in the cytoplasmic fraction. Together with the mapping of 4 distinct bcr related loci our data suggest that the 130 Kda phosphoprotein belongs to a wider family of bcr related gene products.
人类慢性粒细胞白血病的特征是9号和22号染色体之间发生相互易位。这导致c-abl原癌基因从9号染色体转移到22号染色体上的bcr基因中。本研究的目的是对bcr及相关基因产物进行表征。制备了针对细菌表达载体中诱导产生的融合trpE-bcr蛋白的抗体。用代谢性[35S]甲硫氨酸标记的白血病细胞系的单克隆和多克隆抗体进行免疫沉淀，结果显示，在含有bcr-abl融合转录本的费城染色体阳性细胞中存在210、160和130 kDa的蛋白质。在费城染色体阴性细胞中仅存在160和130 kDa的蛋白质。体外激酶分析表明，130 kDa的蛋白质是一种主要在丝氨酸上磷酸化的磷蛋白。部分蛋白酶解表明，p210和p130具有共同的结构域。在亚细胞分级分离实验中，p130与p210 bcr-abl共定位于细胞质部分。结合4个不同的bcr相关基因座的定位，我们的数据表明，130 kDa的磷蛋白属于一个更广泛的bcr相关基因产物家族。
