Brauer Madita, Lassek Christian, Hinze Christian, Hoyer Juliane, Becher Dörte, Jahn Dieter, Sievers Susanne, Riedel Katharina
Department for Microbial Physiology and Molecular Biology, Institute of Microbiology, University of Greifswald, Greifswald, Germany.
Department for Microbial Proteomics, Institute of Microbiology, University of Greifswald, Greifswald, Germany.
Front Microbiol. 2021 Jun 10;12:682111. doi: 10.3389/fmicb.2021.682111. eCollection 2021.
The anaerobic pathogen is perfectly equipped to survive and persist inside the mammalian intestine. When facing unfavorable conditions is able to form highly resistant endospores. Likewise, biofilms are currently discussed as form of persistence. Here a comprehensive proteomics approach was applied to investigate the molecular processes of strain 630Δ underlying biofilm formation. The comparison of the proteome from two different forms of biofilm-like growth, namely aggregate biofilms and colonies on agar plates, revealed major differences in the formation of cell surface proteins, as well as enzymes of its energy and stress metabolism. For instance, while the obtained data suggest that aggregate biofilm cells express both flagella, type IV pili and enzymes required for biosynthesis of cell-surface polysaccharides, the S-layer protein SlpA and most cell wall proteins (CWPs) encoded adjacent to SlpA were detected in significantly lower amounts in aggregate biofilm cells than in colony biofilms. Moreover, the obtained data suggested that aggregate biofilm cells are rather actively growing cells while colony biofilm cells most likely severely suffer from a lack of reductive equivalents what requires induction of the Wood-Ljungdahl pathway and V-type ATPase to maintain cell homeostasis. In agreement with this, aggregate biofilm cells, in contrast to colony biofilm cells, neither induced toxin nor spore production. Finally, the data revealed that the sigma factor SigL/RpoN and its dependent regulators are noticeably induced in aggregate biofilms suggesting an important role of SigL/RpoN in aggregate biofilm formation.
这种厌氧病原体完全有能力在哺乳动物肠道内存活并持续存在。面对不利条件时,它能够形成高度抗性的内生孢子。同样,生物被膜目前也被认为是一种持续存在的形式。在此,我们应用了一种全面的蛋白质组学方法来研究630Δ菌株形成生物被膜的分子过程。对两种不同形式的类生物被膜生长(即聚集生物被膜和琼脂平板上的菌落)的蛋白质组进行比较,揭示了细胞表面蛋白形成以及其能量和应激代谢酶方面的主要差异。例如,虽然获得的数据表明聚集生物被膜细胞表达鞭毛、IV型菌毛以及细胞表面多糖生物合成所需的酶,但在聚集生物被膜细胞中检测到的S层蛋白SlpA和与SlpA相邻编码的大多数细胞壁蛋白(CWPs)的量明显低于菌落生物被膜细胞。此外,获得的数据表明聚集生物被膜细胞是相当活跃生长的细胞,而菌落生物被膜细胞很可能严重缺乏还原当量,这需要诱导伍德-Ljungdahl途径和V型ATP酶来维持细胞内稳态。与此一致的是,与菌落生物被膜细胞相比,聚集生物被膜细胞既不诱导毒素产生也不诱导孢子形成。最后,数据显示聚集生物被膜中σ因子SigL/RpoN及其依赖的调节因子明显被诱导,表明SigL/RpoN在聚集生物被膜形成中起重要作用。