Atarashi K, Kaji A
Department of Microbiology, School of Medicine, University of Pennsylvania, Philadelphia 19104, USA.
J Bacteriol. 2000 Nov;182(21):6154-60. doi: 10.1128/JB.182.21.6154-6160.2000.
Ribosome recycling factor (RRF) of Thermotoga maritima was expressed in Escherichia coli from the cloned T. maritima RRF gene and purified. Expression of T. maritima RRF inhibited growth of the E. coli host in a dose-dependent manner, an effect counteracted by the overexpression of E. coli RRF. T. maritima RRF also inhibited the E. coli RRF reaction in vitro. Genes encoding RRFs from Streptococcus pneumoniae and Helicobacter pylori have been cloned, and they also impair growth of E. coli, although the inhibitory effect of these RRFs was less pronounced than that of T. maritima RRF. The amino acid sequence at positions 57 to 62, 74 to 78, 118 to 122, 154 to 160, and 172 to 176 in T. maritima RRF differed totally from that of E. coli RRF. This suggests that these regions are important for the inhibitory effect of heterologous RRF. We further suggest that bending and stretching of the RRF molecule at the hinge between two domains may be critical for RRF activity and therefore responsible for T. maritima RRF inhibition of the E. coli RRF reaction.
嗜热栖热菌的核糖体循环因子(RRF)通过克隆的嗜热栖热菌RRF基因在大肠杆菌中表达并纯化。嗜热栖热菌RRF的表达以剂量依赖的方式抑制大肠杆菌宿主的生长,这种效应可被大肠杆菌RRF的过表达抵消。嗜热栖热菌RRF在体外也抑制大肠杆菌RRF反应。肺炎链球菌和幽门螺杆菌编码RRF的基因已被克隆,它们也损害大肠杆菌的生长,尽管这些RRF的抑制作用不如嗜热栖热菌RRF明显。嗜热栖热菌RRF中第57至62位、74至78位、118至122位、154至160位和172至176位的氨基酸序列与大肠杆菌RRF完全不同。这表明这些区域对于异源RRF的抑制作用很重要。我们进一步认为,RRF分子在两个结构域之间的铰链处的弯曲和拉伸可能对RRF活性至关重要,因此是嗜热栖热菌RRF抑制大肠杆菌RRF反应的原因。