Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1

MvaT and MvaU are global transcriptional regulators belonging to the H-NS family, and pyocyanin is an important virulence factor produced by Pseudomonas aeruginosa. mvaT mvaU double knockout mutant of P. aeruginosa PAO1 demonstrated pyocyanin abolishment in the previous study. Here, we further explo...

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Autores principales:	Limin Dong, Jing Pang, Xiukun Wang, Youwen Zhang, Guoqing Li, Xinxin Hu, Xinyi Yang, Chung-Dar Lu, Congran Li, Xuefu You
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2020
Materias:	pyocyanin mvat mvau h-ns quorum sensing Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/ce081bb56dc649619eeeb22fbab47e6b
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spelling	oai:doaj.org-article:ce081bb56dc649619eeeb22fbab47e6b2021-11-17T14:21:57ZMechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO12150-55942150-560810.1080/21505594.2019.1708052https://doaj.org/article/ce081bb56dc649619eeeb22fbab47e6b2020-12-01T00:00:00Zhttp://dx.doi.org/10.1080/21505594.2019.1708052https://doaj.org/toc/2150-5594https://doaj.org/toc/2150-5608MvaT and MvaU are global transcriptional regulators belonging to the H-NS family, and pyocyanin is an important virulence factor produced by Pseudomonas aeruginosa. mvaT mvaU double knockout mutant of P. aeruginosa PAO1 demonstrated pyocyanin abolishment in the previous study. Here, we further explored the mechanism. Two main directions were studied: pyocyanin biosynthesis pathway and QS system. The effect on the expression of the pyocyanin biosynthesis genes was evaluated by promoter strength determination and Real-Time PCR assay, and significant changes leading to low pyocyanin production were found. The effect on the QS system was studied by signal molecule quantification using LC-MS/MS and related gene expression measurements using Real-Time PCR. In mvaT mvaU double knockout, the production of 3-oxo-C12-HSL obviously increased, while those of C4-HSL and PQS obviously decreased, and the changes can be recovered by mvaT or mvaU complementation. The expressions of transcriptional activator genes binding with QS system signal molecules were all decreased, resulting in decreased formation of signal-transcriptional activator complexes. And the decreased expression of rhlR and pqsE also led to the lower expression of phzA1 and phzA2. Further exploration found that QS system downregulation may be related to QsrO, a QS system repressor, which was highly upregulated with mvaT mvaU double knockout. Hence, the synthesis of pyocyanin was suffocated and the biofilm formation ability was decreased. These results were also confirmed by transcriptome analysis, which demonstrated similar gene expression changes of the aforementioned genes together with decreased expression of other virulence factor genes regulated by QS system.Limin DongJing PangXiukun WangYouwen ZhangGuoqing LiXinxin HuXinyi YangChung-Dar LuCongran LiXuefu YouTaylor & Francis Grouparticlepyocyaninmvatmvauh-nsquorum sensingInfectious and parasitic diseasesRC109-216ENVirulence, Vol 11, Iss 1, Pp 57-67 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	pyocyanin mvat mvau h-ns quorum sensing Infectious and parasitic diseases RC109-216
spellingShingle	pyocyanin mvat mvau h-ns quorum sensing Infectious and parasitic diseases RC109-216 Limin Dong Jing Pang Xiukun Wang Youwen Zhang Guoqing Li Xinxin Hu Xinyi Yang Chung-Dar Lu Congran Li Xuefu You Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1
description	MvaT and MvaU are global transcriptional regulators belonging to the H-NS family, and pyocyanin is an important virulence factor produced by Pseudomonas aeruginosa. mvaT mvaU double knockout mutant of P. aeruginosa PAO1 demonstrated pyocyanin abolishment in the previous study. Here, we further explored the mechanism. Two main directions were studied: pyocyanin biosynthesis pathway and QS system. The effect on the expression of the pyocyanin biosynthesis genes was evaluated by promoter strength determination and Real-Time PCR assay, and significant changes leading to low pyocyanin production were found. The effect on the QS system was studied by signal molecule quantification using LC-MS/MS and related gene expression measurements using Real-Time PCR. In mvaT mvaU double knockout, the production of 3-oxo-C12-HSL obviously increased, while those of C4-HSL and PQS obviously decreased, and the changes can be recovered by mvaT or mvaU complementation. The expressions of transcriptional activator genes binding with QS system signal molecules were all decreased, resulting in decreased formation of signal-transcriptional activator complexes. And the decreased expression of rhlR and pqsE also led to the lower expression of phzA1 and phzA2. Further exploration found that QS system downregulation may be related to QsrO, a QS system repressor, which was highly upregulated with mvaT mvaU double knockout. Hence, the synthesis of pyocyanin was suffocated and the biofilm formation ability was decreased. These results were also confirmed by transcriptome analysis, which demonstrated similar gene expression changes of the aforementioned genes together with decreased expression of other virulence factor genes regulated by QS system.
format	article
author	Limin Dong Jing Pang Xiukun Wang Youwen Zhang Guoqing Li Xinxin Hu Xinyi Yang Chung-Dar Lu Congran Li Xuefu You
author_facet	Limin Dong Jing Pang Xiukun Wang Youwen Zhang Guoqing Li Xinxin Hu Xinyi Yang Chung-Dar Lu Congran Li Xuefu You
author_sort	Limin Dong
title	Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1
title_short	Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1
title_full	Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1
title_fullStr	Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1
title_full_unstemmed	Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1
title_sort	mechanism of pyocyanin abolishment caused by mvat mvau double knockout in pseudomonas aeruginosa pao1
publisher	Taylor & Francis Group
publishDate	2020
url	https://doaj.org/article/ce081bb56dc649619eeeb22fbab47e6b
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Mechanism of pyocyanin abolishment caused by mvaT mvaU double knockout in Pseudomonas aeruginosa PAO1

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