Caldas T, Binet E, Bouloc P, Richarme G
Biochimie Génétique, Institut Jacques Monod, Université Paris 7,2, place Jussieu, Paris, 75005, France.
Biochem Biophys Res Commun. 2000 May 19;271(3):714-8. doi: 10.1006/bbrc.2000.2702.
We recently identified RrmJ (alias FtsJ), the first encoded protein of the rrmJ-hflB heat shock operon, as an Um(2552) methyltransferase of the 23S rRNA. We now report that the rrmJ-deficient strain exhibits growth and translational defects compared to the wild-type strain. Growth rates of the rrmJ mutant are decreased at both low and high temperatures. Protein synthesis activity is reduced up to 65% when S(30) rrmJ mutant extracts are tested in a coupled in vitro transcription/translation assay. In vitro methylation of these extracts by RrmJ partially restores protein synthesis activity. Polysome profile analysis of the rrmJ strain reveals an increase in the proportion of free 30S and 50S subunits at both 30 and 42 degrees C. These results suggest that the RrmJ-catalyzed methylation of Um(2552) in 23S RNA strengthens ribosomal subunit interactions, increases protein synthesis activity, and improves cell growth rates even at non-heat shock temperatures.
我们最近鉴定出RrmJ(别名FtsJ),它是rrmJ-hflB热休克操纵子的首个编码蛋白,是23S rRNA的Um(2552)甲基转移酶。我们现在报告,与野生型菌株相比,rrmJ缺陷型菌株表现出生长和翻译缺陷。rrmJ突变体在低温和高温下的生长速率均降低。当在体外转录/翻译偶联试验中检测S(30) rrmJ突变体提取物时,蛋白质合成活性降低高达65%。用RrmJ对这些提取物进行体外甲基化可部分恢复蛋白质合成活性。对rrmJ菌株的多核糖体谱分析显示,在30℃和42℃时游离30S和50S亚基的比例均增加。这些结果表明,RrmJ催化的23S RNA中Um(2552)甲基化增强了核糖体亚基相互作用,提高了蛋白质合成活性,甚至在非热休克温度下也能提高细胞生长速率。