Okamoto S, Ochi K
National Food Research Institute, Tsukuba, Ibaraki, Japan.
Mol Microbiol. 1998 Oct;30(1):107-19. doi: 10.1046/j.1365-2958.1998.01042.x.
The Streptomyces coelicolor obg gene, which encodes a putative GTP-binding protein of the Obg/Gtp1 family, was characterized. The obg gene was essential for viability. Introduction of multiple copies of obg into wild-type S. coelicolor suppressed aerial mycelium formation. A single amino acid substitution at any of six positions was introduced into the GTP binding site of Obg, and the mutated proteins were expressed in wild-type cells. Obg(P168-->V) exerted a more accentuated suppressive effect on aerial mycelium formation than did the wild-type Obg protein. In contrast, Obg(G171-->A) accelerated the development of aerial mycelium. These results show that Obg protein functions as a pivotal regulator for the onset of cell differentiation through its ability to bind GTP. Western analysis revealed that expression of obg is regulated in a growth phase-dependent manner, indicating a sharp decrease just after onset of aerial mycelium development or at the end of vegetative growth. Obg was a membrane-bound protein as determined by immunoelectron microscopy.
对天蓝色链霉菌的obg基因进行了表征,该基因编码一种假定的Obg/Gtp1家族的GTP结合蛋白。obg基因对生存能力至关重要。将多个拷贝的obg导入野生型天蓝色链霉菌中会抑制气生菌丝体的形成。在Obg的GTP结合位点的六个位置中的任何一个位置引入单个氨基酸取代,并在野生型细胞中表达突变蛋白。与野生型Obg蛋白相比,Obg(P168-->V)对气生菌丝体形成的抑制作用更为明显。相反,Obg(G171-->A)加速了气生菌丝体的发育。这些结果表明,Obg蛋白通过其结合GTP的能力作为细胞分化起始的关键调节因子发挥作用。蛋白质免疫印迹分析显示,obg的表达以生长阶段依赖性方式受到调节,表明在气生菌丝体发育开始后或营养生长结束时急剧下降。通过免疫电子显微镜确定Obg是一种膜结合蛋白。