van der Westhuizen Wouter André, Theron Chrispian William, Boucher Charlotte Enastacia, Bragg Robert Richard
Department of Microbial, Biochemical and Food Biotechnology, Internal Box 61, University of the Free State, P.O. Box 339, Bloemfontein, 9301, South Africa.
Heliyon. 2019 Jul 1;5(7):e02014. doi: 10.1016/j.heliyon.2019.e02014. eCollection 2019 Jul.
Gram-negative bacteria actively secrete outer membrane vesicles into the surrounding environment and these vesicles have been shown to play various physiological and protective roles such as carrying antibiotic-degrading enzymes and acting as decoys against host defences, therefore promoting the pathogenesis of the bacterium. It has been shown that avian pathogenic species can increase vesicle biosynthesis through the acquisition of the gene but the effect this has on the cell by scavenging outer-membrane associated proteins (OmpA, OmpF) into the vesicles during vesicle release have not yet been investigated.
Relative quantitative real-time PCR data obtained from expressing and non-expressing cells showed that during induction, showed a nearly 2-fold down regulation relative to the non-expressing cells during the entire 24 hours, while was expressed at the same level as the non-expressing cells during the first 8 hours of expression. At 24 hours post- expression, was up-regulated 4-fold.
The regulatory effects of the newly described outer-membrane vesicle biosynthesis-related gene, , on has not previously been investigated. As -induced vesicles contain OmpA and OmpF scavenged from the bacterial outer-membrane, potential regulatory effects on the host was investigated. An increase in expression and an insignificant decrease in expression was observed during induction demonstrating that -related biosynthesis is not related to decreased expression, which is one of the potential mechanisms discussed in literature for biosynthesis. Outer-membrane vesicle biosynthesis during over-expression could potentially be accomplished through a different mechanism(s).
革兰氏阴性菌会主动向周围环境分泌外膜囊泡,这些囊泡已被证明发挥着多种生理和保护作用,例如携带抗生素降解酶以及充当抵御宿主防御的诱饵,从而促进细菌的致病过程。研究表明,禽致病性物种可通过获取该基因来增加囊泡生物合成,但在囊泡释放过程中,通过将外膜相关蛋白(OmpA、OmpF)清除到囊泡中,其对细胞产生的影响尚未得到研究。
从表达和不表达该基因的细胞中获得的相对定量实时PCR数据显示,在诱导过程中,相对于不表达的细胞,在整个24小时内显示出近2倍的下调,而在表达的前8小时内,其表达水平与不表达的细胞相同。在表达后24小时,该基因上调了4倍。
此前尚未研究新描述的外膜囊泡生物合成相关基因对该基因的调控作用。由于该基因诱导产生的囊泡含有从细菌外膜清除的OmpA和OmpF,因此研究了其对宿主的潜在调控作用。在该基因诱导过程中观察到其表达增加,而另一个基因的表达无显著下降,这表明该基因相关的生物合成与文献中讨论的生物合成潜在机制之一——另一个基因表达降低无关。在该基因过表达期间,外膜囊泡生物合成可能通过不同的机制来完成。