Engman Jakob, Negrea Aurel, Sigurlásdóttir Sara, Geörg Miriam, Eriksson Jens, Eriksson Olaspers Sara, Kuwae Asaomi, Sjölinder Hong, Jonsson Ann-Beth
Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden.
Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden
Infect Immun. 2016 Apr 22;84(5):1501-1513. doi: 10.1128/IAI.01463-15. Print 2016 May.
Neisseria meningitidis autoaggregation is an important step during attachment to human cells. Aggregation is mediated by type IV pili and can be modulated by accessory pilus proteins, such as PilX, and posttranslational modifications of the major pilus subunit PilE. The mechanisms underlying the regulation of aggregation remain poorly characterized. Polynucleotide phosphorylase (PNPase) is a 3'-5' exonuclease that is involved in RNA turnover and the regulation of small RNAs. In this study, we biochemically confirm that NMC0710 is the N. meningitidis PNPase, and we characterize its role in N. meningitidis pathogenesis. We show that deletion of the gene encoding PNPase leads to hyperaggregation and increased adhesion to epithelial cells. The aggregation induced was found to be dependent on pili and to be mediated by excessive pilus bundling. PNPase expression was induced following bacterial attachment to human cells. Deletion of PNPase led to global transcriptional changes and the differential regulation of 469 genes. We also demonstrate that PNPase is required for full virulence in an in vivo model of N. meningitidis infection. The present study shows that PNPase negatively affects aggregation, adhesion, and virulence in N. meningitidis.
脑膜炎奈瑟菌的自身聚集是其黏附人类细胞过程中的重要一步。聚集由IV型菌毛介导,可被辅助菌毛蛋白(如PilX)以及主要菌毛亚基PilE的翻译后修饰所调节。聚集调节的潜在机制仍未得到充分表征。多核苷酸磷酸化酶(PNPase)是一种3'-5'核酸外切酶,参与RNA周转和小RNA的调节。在本研究中,我们通过生化方法证实NMC0710是脑膜炎奈瑟菌的PNPase,并对其在脑膜炎奈瑟菌致病机制中的作用进行了表征。我们发现,编码PNPase的基因缺失会导致过度聚集以及对上皮细胞黏附增加。发现诱导的聚集依赖于菌毛,并由过度的菌毛成束介导。细菌黏附人类细胞后会诱导PNPase表达。PNPase缺失导致全局转录变化以及469个基因的差异调节。我们还证明,在脑膜炎奈瑟菌感染的体内模型中,PNPase是完全毒力所必需的。本研究表明,PNPase对脑膜炎奈瑟菌的聚集、黏附和毒力具有负面影响。