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首页> 外文期刊>Molecular and Cellular Biology >Cell-dependent efficiency of reiterated nuclear signals in a mutant simian virus 40 oncoprotein targeted to the nucleus.
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Cell-dependent efficiency of reiterated nuclear signals in a mutant simian virus 40 oncoprotein targeted to the nucleus.

机译：靶向细胞核的突变猿猴病毒40癌蛋白中重申的核信号的细胞依赖性效率。

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摘要

We investigated the requisites for, and functional consequences of, the relocation to the nucleus of a transforming nonkaryophilic mutant of the simian virus 40 large T antigen (a natural deletion mutant lacking an internal large-T-antigen domain that includes the signal for nuclear transport). Synthetic oligonucleotides were used to obtain gene variants with one or more copies of the signal-specifying sequence inserted near the gene 3' end, in a region dispensable for the main large-T-antigen functions. The analysis of stable transfectant populations showed that mouse NIH 3T3 cells, rat embryo fibroblasts, and simian CS cells (a subclone of CV1 cells) differed considerably in their ability to localize some variant molecules into the nucleus. CS cells were always the most efficient, and NIH 3T3 cells were the least efficient. The nuclear localization improved either with reiteration of the signal or with a left-flank modification of the signal amino acid context. Three signals appeared to be necessary and sufficient, even in NIH 3T3 cells, to obtain a nuclear accumulation comparable to that of wild-type simian virus 40 large T antigen; other signal-cell combinations caused a large variability in subcellular localization among cells of the same population, as if the nuclear uptake of some molecules depended on individual cell states. The effect of the modified location on the competence of the protein to alter cell growth was examined by comparing the activity of variants containing either the normal signal or a signal with a mutation (corresponding to large-T-antigen amino acid 128) that prevented nuclear transport. It was found that the nuclear variant was slightly more active than the cytoplasmic variants in rat embryo fibroblasts and NIH 3T3 cells and was notably less active in CS cells.
机译：我们调查了猿猴病毒40大T抗原的转化非亲核突变体（自然缺失突变体缺乏内部大T抗原结构域，其中包括核转运信号）的转化的非亲核突变体重新定位到核中的必要条件和功能后果）。合成的寡核苷酸用于获得基因变异体，该变异体具有一个或多个拷贝的信号指定序列插入基因3'末端附近，该区域可分配给主要的大T抗原功能。稳定转染子群体的分析表明，小鼠NIH 3T3细胞，大鼠胚胎成纤维细胞和猿猴CS细胞（CV1细胞的亚克隆）在将某些变异分子定位到细胞核中的能力差异很大。 CS细胞始终是效率最高的，而NIH 3T3细胞效率最低。通过信号的重复或信号氨基酸上下文的左翼修饰来改善核定位。甚至在NIH 3T3细胞中，三个信号似乎也是必需和充分的，以获得与野生型猿猴病毒40大T抗原相当的核积累。其他信号-细胞组合在同一种群的细胞之间引起亚细胞定位的巨大差异，就好像某些分子的核吸收取决于单个细胞的状态一样。通过比较含有正常信号或具有防止核细胞核抗原突变的信号（对应于大T抗原128）的变异体的活性，研究了修饰位置对蛋白质改变细胞生长能力的影响。运输。发现在大鼠胚胎成纤维细胞和NIH 3T3细胞中，核变体的活性略高于细胞质变体，而在CS细胞中则明显较弱。

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