Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita-shi, Osaka 565-0871, Japan.
J Biosci Bioeng. 2012 Jul;114(1):17-22. doi: 10.1016/j.jbiosc.2012.02.022. Epub 2012 May 16.
Yeast RNA is a good source of nucleotide seasoning, and more than half of yeast RNA consists of ribosomal RNA (rRNA). Previously, we reported the development of a Saccharomyces cerevisiae strain displaying a 1.4- to 2.3-times higher RNA content than the wild-type strain through the isolation of dominant suppressors (designated SupA to SupG strains) from a Δrrn10 disruptant showing decreased rRNA transcription. In the present study, the cloning of one of the genes responsible for the suppression was attempted using a genomic library from the SupD strain. NOP15, a gene involved in ribosome biogenesis, was found to be responsible for suppressing the growth defect of the Δrrn10 disruptant. The isolated NOP15 allele (designated NOP15(T-279C)) possessed a single T to C substitution at nucleotide position-279 of NOP15. The transcription level of NOP15(T-279C) in the originally isolated SupD strain was 2-fold higher than that in the Δrrn10 disruptant. Furthermore, a dose-dependent relationship between the transcription level of NOP15 and total amount of RNA in the Δrrn10 disruptant was observed: the enhanced transcription due to the NOP15(T-279C) allele is involved in the suppression mechanisms in the SupD strain. Introduction of the NOP15(T-279C) allele into the wild-type strain increased the total RNA content by 1.4-fold. These results indicate that the transcription level of NOP15 is an important determinant of the productivity of RNA and that its increased transcription provides an effective approach to obtain higher RNA yields in yeast.
酵母 RNA 是核苷酸调味料的良好来源,其中超过一半的酵母 RNA 由核糖体 RNA(rRNA)组成。之前,我们通过从显示 rRNA 转录减少的Δrrn10 缺失突变体中分离显性抑制子(命名为 SupA 到 SupG 株),报道了一株酿酒酵母菌株的开发,该菌株的 RNA 含量比野生型菌株高 1.4 到 2.3 倍。在本研究中,使用来自 SupD 株的基因组文库尝试克隆负责抑制的一个基因。NOP15 是一个参与核糖体生物发生的基因,被发现负责抑制Δrrn10 缺失突变体的生长缺陷。分离的 NOP15 等位基因(命名为 NOP15(T-279C))在 NOP15 的核苷酸位置-279 处具有单个 T 到 C 取代。最初分离的 SupD 株中 NOP15(T-279C)的转录水平比Δrrn10 缺失突变体高 2 倍。此外,在Δrrn10 缺失突变体中观察到 NOP15 的转录水平与总 RNA 量之间存在剂量依赖性关系:由于 NOP15(T-279C)等位基因导致的增强转录参与了 SupD 株中的抑制机制。将 NOP15(T-279C)等位基因引入野生型菌株可使总 RNA 含量增加 1.4 倍。这些结果表明,NOP15 的转录水平是 RNA 生产力的重要决定因素,其转录水平的增加为在酵母中获得更高的 RNA 产量提供了一种有效方法。