Mutation from guanine to adenine in 25S rRNA at the position equivalent to E-coli A2058 does not confer erythromycin sensitivity in Sacchromyces cerevisae
Mutation from guanine to adenine in 25S rRNA at the position equivalent to E-coli A2058 does not confer erythromycin sensitivity in Sacchromyces cerevisae
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DOI：
10.1261/rna.786408
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发表时间：
2008-03-01
期刊：
影响因子：
4.5
通讯作者：
Zengel, Janice M.
中科院分区：
文献类型：
--
作者：
Bommakanti, Ananth S.;Lindahl, Lasse;Zengel, Janice M.
大环内酯红霉素结合到肽基转移酶中心（PTC）附近的原核核糖体大亚基上，并抑制新肽链延伸超过几个氨基酸。核苷酸A2058和A2059（E.大肠杆菌编号）在红霉素的结合中起关键作用，并且核苷酸A2058突变赋予革兰氏阳性和革兰氏阴性细菌红霉素抗性。在原核和真核核糖体的PTC中存在高度的序列和结构相似性。然而，真核生物核糖体对红霉素具有抗性，并且在与E相当的位置上存在G。大肠杆菌A2058核苷酸被认为是原因。为了验证这一假设，我们在酿酒酵母25S rRNA的这个位置引入了G到A突变，并分析了对红霉素的敏感性。突变酵母核糖体的生长研究和红霉素结合试验均未表明突变酵母菌株对红霉素有任何敏感性。这些结果表明，核苷酸2058的身份不是唯一的决定因素，负责酵母和原核生物之间的红霉素敏感性的差异。
The macrolide erythromycin binds to the large subunit of the prokaryotic ribosome near the peptidyltransferase center (PTC) and inhibits elongation of new peptide chains beyond a few amino acids. Nucleotides A2058 and A2059 (E. coli numbering) in 23S rRNA play a crucial role in the binding of erythromycin, and mutation of nucleotide A2058 confers erythromycin resistance in both Gram-positive and Gram-negative bacteria. There are high levels of sequence and structural similarity in the PTC of prokaryotic and eukaryotic ribosomes. However, eukaryotic ribosomes are resistant to erythromycin and the presence of a G at the position equivalent to E. coli nucleotide A2058 is believed to be the reason. To test this hypothesis, we introduced a G to A mutation at this position of the yeast Saccharomyces cerevisiae 25S rRNA and analyzed sensitivity toward erythromycin. Neither growth studies nor erythromycin binding assays on mutated yeast ribosomes indicated any erythromycin sensitivity in mutated yeast strains. These results suggest that the identity of nucleotide 2058 is not the only determinant responsible for the difference in erythromycin sensitivity between yeast and prokaryotes.