The v-rel oncogene product is complexed to a 40-kDa phosphoprotein in transformed lymphoid cells.
The v-rel oncogene product is complexed to a 40-kDa phosphoprotein in transformed lymphoid cells.
复制标题
v-rel 癌基因产物在转化的淋巴细胞中与 40-kDa 磷蛋白复合。
DOI：
10.1073/pnas.85.8.2479
复制
发表时间：
1988
影响因子：
11.1
通讯作者：
BoseJr,HR
中科院分区：
文献类型：
--
作者：
Tung,HY;Bargmann,WJ;Lim,MY;BoseJr,HR
禽网状内皮增生病毒（REV-T）的v-rel致癌基因编码的转化蛋白是转化细胞细胞质中拷贝数极低的分子。用Sephacryl S-300凝胶过滤法对rev -t转化淋巴细胞株的胞浆提取物进行了分析，发现大部分v-rel致癌基因产物pp59v-rel被洗脱，其表观分子质量为400 kDa。该复合物的大小通过快速蛋白液相色谱凝胶过滤柱的分析得到证实。用Sephacryl S-200进行序次凝胶过滤，用针对pp59v-rel的单克隆抗体进行免疫亲和层析，得到40 kda的细胞蛋白与pp59v-rel共化。从[35S]蛋氨酸标记的细胞提取物中，40-kDa细胞蛋白也可以与pp59v-rel一起免疫沉淀，这表明pp59v-rel在转化的淋巴样细胞中与40-kDa蛋白络合。在体外用[γ - 32p]ATP和10 mM MgCl2孵育含有pp59v-rel的高纯化制剂时，59-和40-kDa蛋白都被磷酸化。然而，在含有pp59v-rel的高纯度制剂中，激酶的身份是未知的。用[32P]正磷酸盐标记的rev -t转化淋巴细胞提取物中回收的免疫复合物也含有59和40 kda的磷酸化蛋白。这些观察结果表明，pp59v-rel在转化淋巴样细胞的细胞质中与40 kda的细胞磷酸化蛋白复合物形成400 kda的异聚物。
The transforming protein encoded by the v-rel oncogene of avian reticuloendotheliosis virus (REV-T) is a very low copy number molecule in the cytosol of transformed cells. Analysis of cytosolic extracts from a REV-T-transformed lymphoid cell line by gel filtration on Sephacryl S-300 indicated that most of the v-rel oncogene product, pp59v-rel, eluted with an apparent molecular mass of 400 kDa. The size of this complex was confirmed by analysis on a fast-protein liquid chromatography gel filtration column. A 40-kDa cellular protein copurified with pp59v-rel on sequential gel filtration on Sephacryl S-200 and immunoaffinity chromatography with a monoclonal antibody directed against pp59v-rel. The 40-kDa cellular protein could also be immunoprecipitated together with pp59v-rel from cell extracts of [35S]methionine-labeled cells, suggesting that pp59v-rel is complexed with the 40-kDa protein in transformed lymphoid cells. Both the 59- and 40-kDa proteins were phosphorylated when the highly purified preparation containing pp59v-rel was incubated with [gamma-32P]ATP and 10 mM MgCl2 in vitro. The identity of the kinase in the highly purified preparation containing pp59v-rel, however, is unknown. Immune complexes recovered from extracts of REV-T-transformed lymphoid cells labeled with [32P]orthophosphate also contained the 59- and 40-kDa phosphoproteins. These observations suggest that pp59v-rel is complexed with a 40-kDa cellular phosphoprotein to form a 400-kDa heteropolymer in the cytoplasm of transformed lymphoid cells.