Mechanisms underlying the virulence regulation of new Vibrio alginolyticus ncRNA Vvrr1 with a comparative proteomic analysis
Mechanisms underlying the virulence regulation of new Vibrio alginolyticus ncRNA Vvrr1 with a comparative proteomic analysis
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新溶藻弧菌 ncRNA Vvrr1 毒力调控机制的比较蛋白质组学分析
DOI：
10.1080/22221751.2019.1687261
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发表时间：
2019-01
影响因子：
13.2
通讯作者：
Huang Lixing
中科院分区：
文献类型：
--
作者：
Zuo Yanfei;Zhao Lingmin;Xu Xiaojin;Zhang Jiaonan;Zhang Jiaolin;Yan Qingpi;Huang Lixing
近年来，受气候变化和海水温度上升的影响，溶藻弧菌感染的发病率有所上升。Vibrio毒力调控RNA 1 （Vvrr1）是一种新发现的非编码RNA (ncRNA)，根据之前的RNA-seq预测其与溶藻弧菌的粘附能力密切相关。本研究通过构建过表达Vvrr1的菌株，利用蛋白质组测序技术，对Vvrr1的靶基因进行了全面筛选和验证。Pyruvate kinase I （pykF）基因是Vvrr1参与毒力调控的主要靶基因。与野生菌株相比，vvrr1过表达菌株和pykf沉默菌株的粘附能力、生物膜形成和毒力均显著降低。与Vvrr1的过表达类似，pykF的沉默也降低了ndk、eno、sdhB、glpF、cysH等毒力基因的表达水平。同时，通过构建“pykF-GFP”融合表达质粒，在大肠杆菌中进行GFP报告基因分析，发现含有Vvrr1全ncRNA序列载体的菌株的荧光强度明显减弱。这表明Vvrr1通过与毒力基因pykF相互作用参与了溶藻弧菌毒力调控机制。
ABSTRACT The incidence of Vibrio alginolyticus infections has increased in recent years due to the influence of climate change and rising sea temperature. Vibrio virulence regulatory RNA 1 (Vvrr1) is a newly found noncoding RNA (ncRNA) predicted to be closely related to the adhesion ability of V. alginolyticus based on the previous RNA-seq. In this study, the target genes of Vvrr1 were fully screened and verified by constructing Vvrr1-overexpressing strains and using the proteome sequencing technology. Pyruvate kinase I (pykF) gene was predicted to be a chief target gene of Vvrr1 involved in virulence regulation. The adhesion ability, biofilm formation and virulence were significantly reduced in the Vvrr1-overexpressing and the pykF-silenced strain compared with the wild strains. Similar to the overexpression of Vvrr1, the silencing of pykF also reduced the expression level of virulence genes, such as ndk, eno, sdhB, glpF, and cysH. Meanwhile, by constructing the “pykF-GFP” fusion expression plasmid and using the GFP reporter gene analysis in Escherichia coli, the fluorescence intensity of the strain containing Vvrr1 whole ncRNA sequence vector was found to be significantly weakened. These indicated that Vvrr1 participated in the virulence regulation mechanism of V. alginolyticus by interacting with the virulence gene pykF.
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影响因子：
4.5
作者：
De Backer S;Sabirova J;De Pauw I;De Greve H;Hernalsteens JP;Goossens H;Malhotra-Kumar S
通讯作者：
Malhotra-Kumar S
影响因子：
6.7
作者：
Hua Yu;X. Rao;Kebin Zhang
通讯作者：
Hua Yu;X. Rao;Kebin Zhang
影响因子：
4.7
作者：
Meimei Zhang;Yingxue Qin;Lixing Huang;Q. Yan;Leilei Mao;Xiaojin Xu;Suyun Wang;Mengmeng Zhang;Liwei Chen
通讯作者：
Meimei Zhang;Yingxue Qin;Lixing Huang;Q. Yan;Leilei Mao;Xiaojin Xu;Suyun Wang;Mengmeng Zhang;Liwei Chen
DOI：
10.1016/j.ijfoodmicro.2018.06.003
发表时间：
2018-10-20
影响因子：
5.4
作者：
Koomen, Jeroen;den Besten, Heidy M. W.;Abee, Tjakko
通讯作者：
Abee, Tjakko
影响因子：
10.7
作者：
Peng F;Widmann S;Wünsche A;Duan K;Donovan KA;Dobson RCJ;Lenski RE;Cooper TF
通讯作者：
Cooper TF