Lu Q, Inouye M
Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
J Bacteriol. 1998 Oct;180(19):5243-6. doi: 10.1128/JB.180.19.5243-5246.1998.
Era, a Ras-like GTP-binding protein in Escherichia coli, has been shown to be essential for growth. However, its cellular functions still remain elusive. In this study, a genetic screening of an E. coli genomic library was performed to identify those genes which can restore the growth ability of a cold-sensitive mutant, Era(Cs) (E200K), at a restrictive temperature when expressed in a multicopy plasmid. Among eight suppressors isolated, six were located at 1 min of the E. coli genomic map, and the gene responsible for the suppression of Era(Cs) (E200K) was identified as the ksgA gene for 16S rRNA transmethylase, whose mutation causes a phenotype of resistance to kasugamycin, a translation initiation inhibitor. This is the first demonstration of suppression of impaired function of Era by overproduction of a functional enzyme. A possible mechanism of the suppression of the Era cold-sensitive phenotype by KsgA overproduction is discussed.
Era是大肠杆菌中一种类Ras的GTP结合蛋白,已被证明对生长至关重要。然而,其细胞功能仍然难以捉摸。在本研究中,对大肠杆菌基因组文库进行了遗传筛选,以鉴定那些在多拷贝质粒中表达时能够在限制温度下恢复冷敏感突变体Era(Cs)(E200K)生长能力的基因。在分离出的八个抑制子中,有六个位于大肠杆菌基因组图谱的1分钟处,负责抑制Era(Cs)(E200K)的基因被鉴定为16S rRNA甲基转移酶的ksgA基因,其突变导致对翻译起始抑制剂卡那霉素产生抗性的表型。这是首次证明通过过量生产功能性酶来抑制Era功能受损。本文还讨论了通过过量生产KsgA抑制Era冷敏感表型的可能机制。