Netterling Sakura, Bäreclev Caroline, Vaitkevicius Karolis, Johansson Jörgen
Department of Molecular Biology, Molecular Infection Medicine, Sweden (MIMS), and Umeå Center for Microbial Research (UCMR), Umeå University, Umeå, Sweden.
Department of Molecular Biology, Molecular Infection Medicine, Sweden (MIMS), and Umeå Center for Microbial Research (UCMR), Umeå University, Umeå, Sweden
Infect Immun. 2015 Oct 19;84(1):67-76. doi: 10.1128/IAI.00849-15. Print 2016 Jan.
RNA helicases have been shown to be important for the function of RNA molecules at several levels, although their putative involvement in microbial pathogenesis has remained elusive. We have previously shown that Listeria monocytogenes DExD-box RNA helicases are important for bacterial growth, motility, ribosomal maturation, and rRNA processing. We assessed the importance of the RNA helicase Lmo0866 (here named CshA) for expression of virulence traits. We observed a reduction in hemolytic activity in a strain lacking CshA compared to the wild type. This phenomenon was less evident in strains lacking other RNA helicases. The reduced hemolysis was accompanied by lower expression of major listerial virulence factors in the ΔcshA strain, mainly listeriolysin O, but also to some degree the actin polymerizing factor ActA. Reduced expression of these virulence factors in the strain lacking CshA did not, however, correlate with a decreased level of the virulence regulator PrfA. When combining the ΔcshA knockout with a mutation creating a constitutively active PrfA protein (PrfA*), the effect of the ΔcshA knockout on LLO expression was negated. These data suggest a role for the RNA helicase CshA in posttranslational activation of PrfA. Surprisingly, although the expression of several virulence factors was reduced, the ΔcshA strain did not demonstrate any reduced ability to infect nonphagocytic cells compared to the wild-type strain.
RNA解旋酶已被证明在多个层面上对RNA分子的功能很重要,尽管它们在微生物致病机制中的假定作用仍不明确。我们之前已经表明,单核细胞增生李斯特菌的DExD-box RNA解旋酶对细菌生长、运动性、核糖体成熟和rRNA加工很重要。我们评估了RNA解旋酶Lmo0866(此处命名为CshA)对毒力特性表达的重要性。我们观察到,与野生型相比,缺乏CshA的菌株溶血活性降低。在缺乏其他RNA解旋酶的菌株中,这种现象不太明显。溶血活性降低伴随着ΔcshA菌株中主要李斯特菌毒力因子的表达降低,主要是溶血素O,但在一定程度上还有肌动蛋白聚合因子ActA。然而,在缺乏CshA的菌株中,这些毒力因子表达的降低与毒力调节因子PrfA水平的降低并无关联。当将ΔcshA基因敲除与产生组成型活性PrfA蛋白(PrfA*)的突变相结合时,ΔcshA基因敲除对LLO表达的影响被消除。这些数据表明RNA解旋酶CshA在PrfA的翻译后激活中发挥作用。令人惊讶的是,尽管几种毒力因子的表达降低了,但与野生型菌株相比,ΔcshA菌株感染非吞噬细胞的能力并未表现出任何降低。