Clegg D O, Koshland D E
Proc Natl Acad Sci U S A. 1984 Aug;81(16):5056-60. doi: 10.1073/pnas.81.16.5056.
A recombinant DNA approach has been used to study intracellular signaling in the bacterial sensing system. The Escherichia coli cheY gene, whose function is unknown, has been subcloned behind the synthetic inducible tac promoter. The resulting plasmid directs the synthesis of the Y protein in response to isopropyl beta-D-thiogalactoside, independent of its usual operon control. When this construct was introduced into wild-type and mutant cells, the Y protein caused a clockwise rotational bias in the flagellar motors. This effect was observed even in heavily biased counterclockwise strains lacking most of the central chemotaxis processing genes. The results show that the Y protein has a direct influence on flagellar rotation not requiring other processing genes of the sensing system. The Y protein appears to bind directly to a part of the flagellar motor, probably the flaA gene product, and it is probably the key element in biasing the motor toward the clockwise rotational direction.
一种重组DNA方法已被用于研究细菌传感系统中的细胞内信号传导。其功能未知的大肠杆菌cheY基因已被亚克隆到合成诱导型tac启动子的下游。所得质粒可响应异丙基β-D-硫代半乳糖苷指导Y蛋白的合成,而与其通常的操纵子控制无关。当将此构建体导入野生型和突变型细胞时,Y蛋白会在鞭毛马达中引起顺时针旋转偏向。即使在缺乏大多数中央趋化性处理基因的严重偏向逆时针旋转的菌株中也观察到了这种效应。结果表明,Y蛋白对鞭毛旋转有直接影响,不需要传感系统的其他处理基因。Y蛋白似乎直接与鞭毛马达的一部分结合,可能是flaA基因产物,并且它可能是使马达偏向顺时针旋转方向的关键元件。