Vimentin phosphorylation by p37mos protein kinase in vitro and generation of a 50-kDa cleavage product in v-mos-transformed cells.
Vimentin phosphorylation by p37mos protein kinase in vitro and generation of a 50-kDa cleavage product in v-mos-transformed cells.
复制标题
体外 p37mos 蛋白激酶对波形蛋白进行磷酸化，并在 v-mos 转化细胞中生成 50 kDa 的裂解产物。
DOI：
10.1016/0042-6822(89)90230-4
复制
发表时间：
1989
期刊：
影响因子：
3.7
通讯作者：
Arlinghaus,RB
中科院分区：
文献类型：
--
作者：
Singh,B;Arlinghaus,RB
先前的研究表明，波形蛋白，一种中间丝蛋白，在急性感染莫洛尼小鼠肉瘤病毒（Mo-MuSV）的细胞中数量减少。在这份报告中，我们提供的证据表明，波形蛋白在Mo-MuSV转化细胞的特异性改变，并证明了p3710 mos蛋白激酶在体外的波形蛋白的特异性磷酸化。磷酸化反应的特异性通过使用由Mo-MuSV和p37 mos在酵母中产生的各种分离物编码的病毒蛋白来证明。缺乏自磷酸化p37 mos能力的磷酸转移结构域突变体不能磷酸化波形蛋白。同样，波形蛋白也没有磷酸化的温度敏感性P85 gag-moskinase来自感染的细胞保持在限制性温度。在ts 110 MuSV转化的NRK细胞中，波形蛋白在转化的允许温度和非允许温度下都被磷酸化。然而，在允许的温度下，检测到波形蛋白（约50 kDa）的改变形式，具有更碱性的等电点和更低的表观分子量。这种50-kDa产物与正常的55-kDa形式的波形蛋白相比高度磷酸化。基于其在二维凝胶中的流动性，50-kDa形式的波形蛋白应该缺少羧基末端。可以想象，这种类型的改变可以调节转化细胞中波形蛋白丝的功能。
Previous studies have shown that vimentin, an intermediate filament protein, is reduced in amount in cells acutely infected with Moloney mouse sarcoma virus (Mo-MuSV). In this report, we provide evidence for specific alteration of vimentin in Mo-MuSV-transformed cells and demonstrate specific phosphorylation of vimentin by the p3710mosprotein kinasein vitro. Specificity of the phosphorylation reaction was demonstrated by using viralmosproteins encoded by various isolates of Mo-MuSV and p37mosproduced in yeast. A phosphotransfer domain mutant lacking the ability to autophosphorylate p37mosfailed to phosphorylate vimentin. Similarly, vimentin was not phosphorylated by the temperature-sensitive P85gag-moskinase derived from infected cells maintained at the restrictive temperature. In tsl 10 MuSV-transformed NRK cells, vimentin was phosphorylated at both the permissive and nonpermissive temperatures for transformation. However, at the permissive temperature, an altered form of vimentin (about 50 kDa) with a more basic isoelectric point and lower apparent molecular weight was detected. This 50-kDa product was highly phosphorylated as compared to the bulk of the normal 55-kDa form of vimentin. On the basis of its mobility in two-dimensional gels, the 50-kDa form of vimentin should lack the carboxy terminus. This type of alteration could conceivably modulate the function of vimentin filaments in the transformed cell.