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首页> 外文期刊>Molecular and Cellular Biology >Ribosomal protein S14 is altered by two-step emetine resistance mutations in Chinese hamster cells.
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Ribosomal protein S14 is altered by two-step emetine resistance mutations in Chinese hamster cells.

机译：核糖体蛋白S14通过中国仓鼠细胞中的两步美因丁抗性突变而改变。

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Four two-dimensional polyacrylamide gel electrophoresis systems were used to identify 78 Chinese hamster cell ribosomal proteins by the uniform nomenclature based on rat liver ribosomal proteins. The 40S ribosomal subunit protein affected by Chinese hamster ovary (CHO) cell one-step emetine resistance mutations is designated S14 in the standard nomenclature. To seek unambiguous genetic evidence for a cause and effect relationship between CHO cell emetine resistance and mutations in the S14 gene, we mutagenized a one-step CHO cell mutant and isolated second-step mutant clones resistant to 10-fold-higher concentrations of emetine. All of the highly resistant, two-step CHO cell mutants obtained displayed additional alterations in ribosomal protein S14. Hybridization complementation tests revealed that the two-step CHO cell emetine resistance mutants were members of the same complementation group defined by one-step CHO cell mutants, EmtB. Two-step mutants obtained from a Chinese hamster lung cell emetine-resistant clone belong to the EmtA complementation group. The two-step and EmtB mutants elaborated 40S ribosomal subunits, which dissociated to 32S and 40S core particles in buffers containing 0.5 M KCl at 4 degrees C. In contrast, 40S ribosomal subunits purified from all EmtA, one-step EmtB EmtC mutants, and wild-type CHO and lung cells were stable at this temperature in buffers containing substantially higher concentrations of salt. Thus, two-step emtB mutations affect the structure of S14 protein directly and the stability of the 40S ribosomal subunit indirectly.
机译：通过基于大鼠肝核糖体蛋白的统一命名法，使用四个二维聚丙烯酰胺凝胶电泳系统鉴定了78种中国仓鼠细胞核糖体蛋白。在标准命名法中，受中国仓鼠卵巢（CHO）细胞一步艾美汀耐药性突变影响的40S核糖体亚基蛋白被指定为S14。为了寻找明确的遗传学证据，以了解CHO细胞的Emetine耐药性与S14基因突变之间的因果关系，我们诱变了一步CHO细胞突变体，并分离了对Emetine浓度高10倍的第二步突变体克隆。获得的所有高抗性，两步CHO细胞突变体均显示出核糖体蛋白S14的其他变化。杂交互补测试表明，两步CHO细胞的metetine抗性突变体是由一步一步CHO细胞突变体EmtB定义的同一互补组的成员。从中国仓鼠肺细胞曲美汀抗性克隆中获得的两步突变体属于EmtA互补组。两步突变体和EmtB突变体精心构建了40S核糖体亚基，在4°C下与含有0.5 M KCl的缓冲液中的32S和40S核心颗粒解离。相反，从所有EmtA，一步一步EmtB EmtC突变体和在此温度下，野生型CHO和肺细胞在含有实质上较高浓度盐的缓冲液中稳定。因此，两步emtB突变直接影响S14蛋白的结构，并间接影响40S核糖体亚基的稳定性。

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