Department of Molecular Biology, Umeå University, Umeå, Sweden.
J Bacteriol. 2012 Aug;194(16):4377-85. doi: 10.1128/JB.00348-12. Epub 2012 Jun 15.
Listeria monocytogenes, a Gram-positive food-borne human pathogen, is able to grow at temperatures close to 0°C and is thus of great concern for the food industry. In this work, we investigated the physiological role of one DExD-box RNA helicase in Listeria monocytogenes. The RNA helicase Lmo1722 was required for optimal growth at low temperatures, whereas it was dispensable at 37°C. A Δlmo1722 strain was less motile due to downregulation of the major subunit of the flagellum, FlaA, caused by decreased flaA expression. By ribosomal fractionation experiments, it was observed that Lmo1722 was mainly associated with the 50S subunit of the ribosome. Absence of Lmo1722 decreased the fraction of 50S ribosomal subunits and mature 70S ribosomes and affected the processing of the 23S precursor rRNA. The ribosomal profile could be restored to wild-type levels in a Δlmo1722 strain expressing Lmo1722. Interestingly, the C-terminal part of Lmo1722 was redundant for low-temperature growth, motility, 23S rRNA processing, and appropriate ribosomal maturation. However, Lmo1722 lacking the C terminus showed a reduced affinity for the 50S and 70S fractions, suggesting that the C terminus is important for proper guidance of Lmo1722 to the 50S subunit. Taken together, our results show that the Listeria RNA helicase Lmo1722 is essential for growth at low temperatures, motility, and rRNA processing and is important for ribosomal maturation, being associated mainly with the 50S subunit of the ribosome.
李斯特菌是一种革兰氏阳性食源性病原体,能够在接近 0°C 的温度下生长,因此对食品工业非常关注。在这项工作中,我们研究了李斯特菌中一种 DExD-box RNA 解旋酶的生理作用。RNA 解旋酶 Lmo1722 是在低温下最佳生长所必需的,而在 37°C 时则是可有可无的。由于鞭毛的主要亚基 FlaA 的表达下调,导致鞭毛运动减少,Δlmo1722 菌株的运动能力下降。通过核糖体分级分离实验观察到,Lmo1722 主要与核糖体的 50S 亚基相关。Lmo1722 的缺失降低了 50S 核糖体亚基和成熟的 70S 核糖体的比例,并影响了 23S 前体 rRNA 的加工。在 Δlmo1722 菌株中表达 Lmo1722 可以使核糖体图谱恢复到野生型水平。有趣的是,Lmo1722 的 C 端对于低温生长、运动、23S rRNA 加工和适当的核糖体成熟是冗余的。然而,缺乏 C 端的 Lmo1722 对 50S 和 70S 分数的亲和力降低,表明 C 端对于 Lmo1722 正确引导到 50S 亚基是很重要的。总之,我们的研究结果表明,李斯特菌 RNA 解旋酶 Lmo1722 是低温生长、运动和 rRNA 加工所必需的,对核糖体成熟也很重要,主要与核糖体的 50S 亚基相关。