Physiological consequences of inactivation of lgmB and lpxL1, two genes involved in lipid A synthesis in Bordetella bronchiseptica

IF 2.5 4区 生物学 Q3 MICROBIOLOGY
Jesús Pérez-Ortega , Roel M. van Harten , Henk P. Haagsman , Jan Tommassen
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Abstract

To develop a Bordetella bronchiseptica vaccine with reduced endotoxicity, we previously inactivated lpxL1, the gene encoding the enzyme that incorporates a secondary 2-hydroxy-laurate in lipid A. The mutant showed a myriad of phenotypes. Structural analysis showed the expected loss of the acyl chain but also of glucosamine (GlcN) substituents, which decorate the phosphates in lipid A. To determine which structural change causes the various phenotypes, we inactivated here lgmB, which encodes the GlcN transferase, and lpxL1 in an isogenic background and compared the phenotypes. Like the lpxL1 mutation, the lgmB mutation resulted in reduced potency to activate human TLR4 and to infect macrophages and in increased susceptibility to polymyxin B. These phenotypes are therefore related to the loss of GlcN decorations. The lpxL1 mutation had a stronger effect on hTLR4 activation and additionally resulted in reduced murine TLR4 activation, surface hydrophobicity, and biofilm formation, and in a fortified outer membrane as evidenced by increased resistance to several antimicrobials. These phenotypes, therefore, appear to be related to the loss of the acyl chain. Moreover, we determined the virulence of the mutants in the Galleria mellonella infection model and observed reduced virulence of the lpxL1 mutant but not of the lgmB mutant.

lgmB和lpxL1失活的生理后果,这两个基因参与了支气管杆菌脂质A的合成
为了开发一种内毒素性降低的支气管败血博德特氏菌疫苗,我们之前灭活了lpxL1,该基因编码在脂质a中结合次级2-羟基-尿酸盐的酶。该突变体表现出多种表型。结构分析显示了酰基链的预期损失,但也显示了修饰脂质A中磷酸盐的葡糖胺(GlcN)取代基的预期损失。为了确定哪种结构变化导致了各种表型,我们在同源背景下灭活了编码GlcN转移酶的lgmB和lpxL1,并比较了表型。与lpxL1突变一样,lgmB突变导致激活人TLR4和感染巨噬细胞的效力降低,并增加对多粘菌素B的易感性。因此,这些表型与GlcN修饰的丧失有关。lpxL1突变对hTLR4的激活具有更强的影响,此外还导致小鼠TLR4的活化、表面疏水性和生物膜形成减少,并在强化的外膜中,对几种抗菌药物的耐药性增加证明了这一点。因此,这些表型似乎与酰基链的缺失有关。此外,我们在意大利黑加仑菌感染模型中测定了突变体的毒力,并观察到lpxL1突变体的毒力降低,但lgmB突变体的毒力没有降低。
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来源期刊
Research in microbiology
Research in microbiology 生物-微生物学
CiteScore
4.10
自引率
3.80%
发文量
54
审稿时长
16 days
期刊介绍: Research in Microbiology is the direct descendant of the original Pasteur periodical entitled Annales de l''Institut Pasteur, created in 1887 by Emile Duclaux under the patronage of Louis Pasteur. The Editorial Committee included Chamberland, Grancher, Nocard, Roux and Straus, and the first issue began with Louis Pasteur''s "Lettre sur la Rage" which clearly defines the spirit of the journal:"You have informed me, my dear Duclaux, that you intend to start a monthly collection of articles entitled "Annales de l''Institut Pasteur". You will be rendering a service that will be appreciated by the ever increasing number of young scientists who are attracted to microbiological studies. In your Annales, our laboratory research will of course occupy a central position, but the work from outside groups that you intend to publish will be a source of competitive stimulation for all of us."That first volume included 53 articles as well as critical reviews and book reviews. From that time on, the Annales appeared regularly every month, without interruption, even during the two world wars. Although the journal has undergone many changes over the past 100 years (in the title, the format, the language) reflecting the evolution in scientific publishing, it has consistently maintained the Pasteur tradition by publishing original reports on all aspects of microbiology.
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