Ohashi Yoshiaki, Inaoka Takashi, Kasai Koji, Ito Yasuhiro, Okamoto Susumu, Satsu Hideo, Tozawa Yuzuru, Kawamura Fujio, Ochi Kozo
National Food Research Institute, Tsukuba, Ibaraki, Japan.
Biosci Biotechnol Biochem. 2003 Oct;67(10):2245-53. doi: 10.1271/bbb.67.2245.
A DNA microarray technique was used to demonstrate global changes in the transcription pattern of translation-associated genes that encode fifty-four ribosomal proteins including a putative ribosomal gene, and eleven translation factors in sporulating B. subtilis. We found that the mRNA levels of nine genes involved in the translation system, which include the genes for three ribosomal proteins (rpmA, rpmGB, and ctc) and two translation factors (efp, and frr), were maintained at a high level at the onset of sporulation. The ypfD gene, which encodes the ribosomal protein S1 homologue, was also found to be expressed significantly during the early sporulation stage. In order to demonstrate the significance of these genes for sporulation, mutants were constructed using the pMutinT3 disruption vector. We detected an impaired sporulation in the mutants of rpmA (gene for the ribosomal protein L27), efp (elongation factor P), frr (ribosome recycling factor), and ypfD. The effect was especially pronounced in the efp mutant, sporulation of which was entirely abolished without affecting growth. The reduced expression of rpmGB (ribosomal protein L33) resulted in an impaired sporulation only at a high temperature (47 degrees C). Only the rplI mutant, which encodes the ribosomal protein L9, could not be obtained, implying that its function is essential for viability. Thus, we successfully demonstrated the significance of several translation-associated genes in sporulation by using the results of the gene expression profiling.
采用DNA微阵列技术来展示枯草芽孢杆菌芽孢形成过程中,与翻译相关基因转录模式的整体变化,这些基因编码54种核糖体蛋白(包括一个假定的核糖体基因)以及11种翻译因子。我们发现,参与翻译系统的9个基因的mRNA水平,在芽孢形成开始时维持在较高水平,其中包括3种核糖体蛋白(rpmA、rpmGB和ctc)以及2种翻译因子(efp和frr)的基因。还发现,编码核糖体蛋白S1同源物的ypfD基因在芽孢形成早期也有显著表达。为了证明这些基因对芽孢形成的重要性,使用pMutinT3破坏载体构建了突变体。我们检测到rpmA(核糖体蛋白L27基因)、efp(延伸因子P)、frr(核糖体循环因子)和ypfD突变体的芽孢形成受损。这种影响在efp突变体中尤为明显,其芽孢形成完全被阻断,且不影响生长。rpmGB(核糖体蛋白L33)表达的降低仅在高温(47摄氏度)下导致芽孢形成受损。只有编码核糖体蛋白L9的rplI突变体无法获得,这意味着其功能对生存力至关重要。因此,我们通过基因表达谱分析结果成功证明了几个与翻译相关的基因在芽孢形成中的重要性。