Kuwana Ritsuko, Takamatsu Hiromu, Watabe Kazuhito
Faculty of Pharmaceutical Sciences, Setsunan University, Hirakata, Osaka 573-0101, Japan.
J Biochem. 2007 Dec;142(6):681-9. doi: 10.1093/jb/mvm179. Epub 2007 Sep 28.
YxeE is a spore coat protein of Bacillus subtilis. We have previously reported that YxeE can be extracted from yabG mutant spores. In the present study, we analysed the expression and localization of YxeE. Northern analysis detected the transcript of yxeE from sporulating cells 4 h after the onset of sporulation, and revealed that the synthesis of yxeE mRNA was dependent on expression of the SigK RNA polymerase and the GerE regulator. Immunoblotting with anti-YxeE antiserum detected YxeE from sporulating cells 4 h after the onset of sporulation. YxeE was detected in the extracts from mature spores of yabG mutant strain but not in those from wild-type spores. On the other hand, fluorescence microscope observations showed that YxeE-green fluorescent protein (GFP) is located at the surface of both wild-type and yabG spores. Immunoblot analysis with anti-GFP antiserum showed that YxeE-GFP was not digested in yabG-disrupted strain and that only the GFP portion remained in the wild-type yabG background. We conclude that YxeE is a substrate of YabG protease.
YxeE是枯草芽孢杆菌的一种芽孢衣蛋白。我们之前报道过YxeE可以从yabG突变体芽孢中提取出来。在本研究中,我们分析了YxeE的表达和定位。Northern分析在芽孢形成开始4小时后从芽孢形成细胞中检测到yxeE的转录本,并表明yxeE mRNA的合成依赖于SigK RNA聚合酶和GerE调节因子的表达。用抗YxeE抗血清进行免疫印迹分析,在芽孢形成开始4小时后从芽孢形成细胞中检测到YxeE。在yabG突变株的成熟芽孢提取物中检测到YxeE,但在野生型芽孢提取物中未检测到。另一方面,荧光显微镜观察表明,YxeE-绿色荧光蛋白(GFP)位于野生型和yabG芽孢的表面。用抗GFP抗血清进行免疫印迹分析表明,YxeE-GFP在yabG缺失菌株中未被消化,并且在野生型yabG背景中仅保留GFP部分。我们得出结论,YxeE是YabG蛋白酶的底物。