An Ruisheng, Sreevatsan Srinand, Grewal Parwinder S
Entomology Department, The Ohio State University, Wooster, OH 44691, USA.
BMC Microbiol. 2008 Jan 28;8:19. doi: 10.1186/1471-2180-8-19.
The bacterium Moraxella osloensis is a mutualistic symbiont of the slug-parasitic nematode Phasmarhabditis hermaphrodita. In nature, P. hermaphrodita vectors M. osloensis into the shell cavity of the slug host Deroceras reticulatum in which the bacteria multiply and kill the slug. As M. osloensis is the main killing agent, genes expressed by M. osloensis in the slug are likely to play important roles in virulence. Studies on pathogenic interactions between bacteria and lower order hosts are few, but such studies have the potential to shed light on the evolution of bacterial virulence. Therefore, we investigated such an interaction by determining gene expression of M. osloensis in its slug host D. reticulatum by selectively capturing transcribed sequences.
Thirteen M. osloensis genes were identified to be up-regulated post infection in D. reticulatum. Compared to the in vitro expressed genes in the stationary phase, we found that genes of ubiquinone synthetase (ubiS) and acyl-coA synthetase (acs) were up-regulated in both D. reticulatum and stationary phase in vitro cultures, but the remaining 11 genes were exclusively expressed in D. reticulatum and are hence infection specific. Mutational analysis on genes of protein-disulfide isomerase (dsbC) and ubiS showed that the virulence of both mutants to slugs was markedly reduced and could be complemented. Further, compared to the growth rate of wild-type M. osloensis, the dsbC and ubiS mutants showed normal and reduced growth rate in vitro, respectively.
We conclude that 11 out of the 13 up-regulated M. osloensis genes are infection specific. Distribution of these identified genes in various bacterial pathogens indicates that the virulence genes are conserved among different pathogen-host interactions. Mutagenesis, growth rate and virulence bioassays further confirmed that ubiS and dsbC genes play important roles in M. osloensis survival and virulence, respectively in D. reticulatum.
奥斯陆莫拉菌是寄生蛞蝓的线虫——雌雄同体拟小杆线虫的一种互利共生菌。在自然界中,雌雄同体拟小杆线虫将奥斯陆莫拉菌传播到蛞蝓宿主网状细胸蛞蝓的壳腔内,细菌在其中繁殖并杀死蛞蝓。由于奥斯陆莫拉菌是主要的致死因子,该菌在蛞蝓体内表达的基因可能在毒力方面发挥重要作用。关于细菌与低等宿主之间致病相互作用的研究较少,但此类研究有可能揭示细菌毒力的进化。因此,我们通过选择性捕获转录序列来测定奥斯陆莫拉菌在其蛞蝓宿主网状细胸蛞蝓中的基因表达,从而研究这种相互作用。
鉴定出13个奥斯陆莫拉菌基因在感染网状细胸蛞蝓后上调。与体外稳定期表达的基因相比,我们发现泛醌合成酶(ubiS)和酰基辅酶A合成酶(acs)的基因在网状细胸蛞蝓和体外稳定期培养物中均上调,但其余11个基因仅在网状细胸蛞蝓中表达,因此是感染特异性的。对蛋白质二硫键异构酶(dsbC)和ubiS基因的突变分析表明,两个突变体对蛞蝓的毒力均显著降低,且可互补。此外,与野生型奥斯陆莫拉菌的生长速率相比,dsbC和ubiS突变体在体外分别表现出正常和降低的生长速率。
我们得出结论,上调的13个奥斯陆莫拉菌基因中有11个是感染特异性的。这些已鉴定基因在各种细菌病原体中的分布表明,毒力基因在不同病原体 - 宿主相互作用中是保守的。诱变、生长速率和毒力生物测定进一步证实,ubiS和dsbC基因分别在奥斯陆莫拉菌在网状细胸蛞蝓中的存活和毒力方面发挥重要作用。