Nouri N, Fallon A M
In Vitro Cell Dev Biol. 1987 Mar;23(3):175-80. doi: 10.1007/BF02623577.
Somatic cell mutants resistant to drugs that interact with the eukaryotic ribosome provide a useful tool for studies on ribosome structure, function, and genetics. From Aedes albopictus (mosquito) cells, cycloheximide-resistant mutants (Cx-705 and Cx-738) that were about 30-fold more resistant to cycloheximide than the parental cells have been obtained. The observation that protein synthesis in cell-free lysates from Cx-705 and Cx-738 cells was resistant to cycloheximide led us to suspect that the alteration in these mutants might affect the ribosome. The present studies show that the cycloheximide-resistant cells grow poorly and eventually die at 34.5 degrees C, a temperature at which wild-type cells grow normally. Relative to control cells, the cycloheximide-resistant cells show increased sensitivity to G-418, another antibiotic that interacts with the eukaryotic ribosome. However, there were no differences between cycloheximide-resistant cells and wild-type cells in sensitivity to puromycin, emetine, or cryptopleurine. Cx-705 cells were predominantly diploid; in contrast, the frequency of tetraploid nuclei in Cx-738 cells was about 40%.
对与真核核糖体相互作用的药物具有抗性的体细胞突变体为核糖体结构、功能及遗传学研究提供了有用的工具。从白纹伊蚊(蚊子)细胞中已获得对放线菌酮具有抗性的突变体(Cx - 705和Cx - 738),它们对放线菌酮的抗性比亲代细胞高约30倍。对Cx - 705和Cx - 738细胞无细胞裂解物中蛋白质合成对放线菌酮具有抗性的观察使我们怀疑这些突变体中的改变可能会影响核糖体。目前的研究表明,对放线菌酮具有抗性的细胞生长不良,并最终在34.5摄氏度下死亡,而野生型细胞在该温度下能正常生长。相对于对照细胞,对放线菌酮具有抗性的细胞对另一种与真核核糖体相互作用的抗生素G - 418表现出更高的敏感性。然而,对放线菌酮具有抗性的细胞与野生型细胞在对嘌呤霉素、依米丁或隐耳草碱的敏感性方面没有差异。Cx - 705细胞主要是二倍体;相比之下,Cx - 738细胞中四倍体细胞核的频率约为40%。