Loh Pek Chin, Morimoto Takuya, Matsuo Yoshitaka, Oshima Taku, Ogasawara Naotake
Graduate School of Information Science, Nara Institute of Science and Technology, Japan.
Genes Genet Syst. 2007 Aug;82(4):281-9. doi: 10.1266/ggs.82.281.
Bacterial genome sequencing has revealed a novel family of P-loop GTPases that are often essential for growth. Accumulating evidence suggests that these proteins are involved in biogenesis of the 30S or 50S ribosomal subunits. YqeH is a member of this Obg/Era GTPase family, with its function remains to be uncovered. Here, we present results showing that YqeH is involved in the 30S subunit biogenesis in Bacillus subtilis. We observed a reduction in the 70S ribosome and accumulation of the free 50S subunit in YqeH-depleted cells. Interestingly, no free 30S subunit accumulation was evident. Consistent with the theory that YqeH is involved in 30S subunit biogenesis, a precursor of 16S rRNA and its degradation products were detected. Additionally, the reduction of free 30S subunit was not observed in Era-depleted cells. YqeH overexpression did not compensate for growth defects in mutants devoid of Era and vice versa. Moreover, in vitro GTPase analyses showed that YqeH possessed high intrinsic GTPase activity. In contrast, Era showed slow GTPase activity, which was enhanced by the 30S ribosomal subunit. Our findings strongly suggest that YqeH and Era function at distinct checkpoints during 30S subunit assembly. B. subtilis yqeH is classified as an essential gene due to the inability of the IPTG-dependent P(spac)-yqeH mutant to grow on LB or PAB agar plates in the absence of IPTG. However, in our experiments, the P(spac)-yqeH mutant grew in PAB liquid medium without IPTG supplementation, albeit at an impaired rate. This finding raises the interesting possibility that YqeH participates in assembly of the 30S ribosomal subunit as well as other cellular functions essential for growth on solid media.
细菌基因组测序揭示了一个新的P环GTP酶家族,这些酶通常对生长至关重要。越来越多的证据表明,这些蛋白质参与30S或50S核糖体亚基的生物合成。YqeH是这个Obg/Era GTP酶家族的成员,其功能尚待揭示。在这里,我们展示的结果表明YqeH参与枯草芽孢杆菌中30S亚基的生物合成。我们观察到在缺乏YqeH的细胞中70S核糖体减少,游离50S亚基积累。有趣的是,没有明显的游离30S亚基积累。与YqeH参与30S亚基生物合成的理论一致,检测到了16S rRNA的前体及其降解产物。此外,在缺乏Era的细胞中未观察到游离30S亚基的减少。YqeH的过表达不能补偿缺乏Era的突变体中的生长缺陷,反之亦然。此外,体外GTP酶分析表明YqeH具有高内在GTP酶活性。相比之下,Era显示出缓慢的GTP酶活性,30S核糖体亚基可增强这种活性。我们的发现强烈表明YqeH和Era在30S亚基组装过程中的不同检查点发挥作用。枯草芽孢杆菌yqeH由于IPTG依赖性P(spac)-yqeH突变体在没有IPTG的情况下无法在LB或PAB琼脂平板上生长而被归类为必需基因。然而,在我们的实验中,P(spac)-yqeH突变体在没有补充IPTG的PAB液体培养基中生长,尽管生长速度受损。这一发现提出了一个有趣的可能性,即YqeH参与30S核糖体亚基的组装以及在固体培养基上生长所必需的其他细胞功能。