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李斯特菌溶血素功能与生物膜形成有关:转录组学分析。

Hemolysin function of Listeria is related to biofilm formation: transcriptomics analysis.

机构信息

Department of Public Health Laboratory Sciences, West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu, 610061, China.

Shen Zhen Biomed Alliance Biotech Group Co., Ltd, Shenzhen, 518057, China.

出版信息

Vet Res. 2022 Dec 31;53(1):113. doi: 10.1186/s13567-022-01124-y.

Abstract

Listeriolysin O (LLO) is the main virulence protein of Listeria monocytogenes (LM), that helps LM escape lysosomes. We previously found that the cellular immune response elicited by L.ivanovii (LI) is weaker than that elicited by LM. We speculated that this may be related to the function of ivanolysin O (ILO). Here, we constructed hemolysin gene deletion strain, LIΔilo, and a modified strain, LIΔilo::hly, in which ilo was replaced by hly. Prokaryotic transcriptome sequencing was performed on LI, LIΔilo, and LIΔilo::hly. Transcriptome differences between the three strains were compared, and genes and pathways with significant differences between the three strains were analyzed. Prokaryotic transcriptome sequencing results revealed the relationship of ilo to the ribosome, quorum sensing, and phosphotransferase system (PTS) pathways, etc. LIΔilo exhibited attenuated biofilm formation ability compared to LI. Biofilm formation was significantly recovered or even increased after replenishing hly. After knocking out ilo, the relative expression levels of some virulence genes, including sigB, prfA, actA, smcL, and virR, were up-regulated compared to LI. After replenishing hly, these genes were down-regulated compared to LIΔilo. The trend and degree of such variation were not completely consistent when cultured in media containing only monosaccharides or disaccharides. The results confirmed that hemolysin is related to some important biological properties of Listeria, including biofilm formation and virulence gene expression levels. This is the first comprehensive study on ILO function at the transcriptomic level and the first evidence of a relationship between Listeria hemolysin and biofilm formation.

摘要

李斯特菌溶血素 O(LLO)是李斯特菌(LM)的主要毒力蛋白,有助于 LM 逃避溶酶体。我们之前发现李斯特菌(LI)引起的细胞免疫反应比 LM 引起的弱。我们推测这可能与伊瓦诺ysin O(ILO)的功能有关。在此,我们构建了溶血素基因缺失株 LIΔilo 和修饰株 LIΔilo::hly,其中 ilo 被 hly 取代。对 LI、LIΔilo 和 LIΔilo::hly 进行了原核转录组测序。比较了三株之间的转录组差异,并分析了三株之间差异显著的基因和通路。原核转录组测序结果揭示了 ilo 与核糖体、群体感应和磷酸转移酶系统(PTS)途径等的关系。LIΔilo 的生物膜形成能力比 LI 减弱。补充 hly 后,生物膜形成能力明显恢复甚至增加。敲除 ilo 后,一些毒力基因(包括 sigB、prfA、actA、smcL 和 virR)的相对表达水平与 LI 相比上调。补充 hly 后,与 LIΔilo 相比,这些基因下调。在仅含有单糖或二糖的培养基中培养时,这种变化的趋势和程度并不完全一致。结果证实溶血素与李斯特菌的一些重要生物学特性有关,包括生物膜形成和毒力基因表达水平。这是首次在转录组水平上对 ILO 功能进行全面研究,也是首次证明李斯特菌溶血素与生物膜形成之间存在关系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/966f/9805692/c59b1a79d8a8/13567_2022_1124_Fig1_HTML.jpg

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