PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1

Pseudomonas lipases are well-studied, but few studies have examined the mechanisms of lipase expression regulation. As a global regulatory protein, PmrA controls the expression of multiple genes such as the Dot/Icm apparatus, eukaryotic-like proteins, and secreted effectors. In this study, the effec...

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Main Authors: Wu Liu, Menggang Li, Liangcheng Jiao, Pengbo Wang, Yunjun Yan
Format: Article
Language:English
Published: Frontiers Media S.A. 2018-01-01
Series:Frontiers in Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/article/10.3389/fmicb.2017.02690/full
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spelling doaj-ae57b2498bfd4bb6a21bbc81b27680e12020-11-24T21:47:49ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-01-01810.3389/fmicb.2017.02690320347PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1Wu Liu0Menggang Li1Liangcheng Jiao2Pengbo Wang3Yunjun Yan4College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, ChinaCollege of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, ChinaCollege of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, ChinaCollege of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, ChinaKey Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, ChinaPseudomonas lipases are well-studied, but few studies have examined the mechanisms of lipase expression regulation. As a global regulatory protein, PmrA controls the expression of multiple genes such as the Dot/Icm apparatus, eukaryotic-like proteins, and secreted effectors. In this study, the effect of PmrA on expression of the lipase lipA in Pseudomonas aeruginosa PAO1 was investigated by knocking out or overexpressing pmrA, rsmY, and rsmA. PmrA regulated the expression of lipA at both the transcriptional and translational level although translation was the pivotal regulatory mechanism for lipA expression. PmrA also regulated the expression of rsmY. Using gel mobility shift assay and pmrA/rsmY double gene knock-out model, we showed that PmrA directly bound to the promoter sequence of rsmY to regulate lipA expression. Translation of lipA was activated by the PmrA/PmrB system via RsmA. Specifically, the Shine-Dalgarno (SD) sequence located at lipA mRNA was overlapped through combination between RsmA and the AGAUGA sequence, subsequently blocking the 30S ribosomal subunit to the SD sequence, leading to translational inhibition of lipA. Transcriptional repression of RsmY initiated translation of lipA through negative translational regulation of rsmA. In conclusion, this study demonstrated that in P. aeruginosa PAO1, PmrA mainly regulated rsmY expression at a translational level to influence lipA expression. RsmY primarily activated lipA translation via negative translational regulation of rsmA.http://journal.frontiersin.org/article/10.3389/fmicb.2017.02690/fulllipasePseudomonas aeruginosamutationexpressiongene regulation
collection DOAJ
language English
format Article
sources DOAJ
author Wu Liu
Menggang Li
Liangcheng Jiao
Pengbo Wang
Yunjun Yan
spellingShingle Wu Liu
Menggang Li
Liangcheng Jiao
Pengbo Wang
Yunjun Yan
PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1
Frontiers in Microbiology
lipase
Pseudomonas aeruginosa
mutation
expression
gene regulation
author_facet Wu Liu
Menggang Li
Liangcheng Jiao
Pengbo Wang
Yunjun Yan
author_sort Wu Liu
title PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1
title_short PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1
title_full PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1
title_fullStr PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1
title_full_unstemmed PmrA/PmrB Two-Component System Regulation of lipA Expression in Pseudomonas aeruginosa PAO1
title_sort pmra/pmrb two-component system regulation of lipa expression in pseudomonas aeruginosa pao1
publisher Frontiers Media S.A.
series Frontiers in Microbiology
issn 1664-302X
publishDate 2018-01-01
description Pseudomonas lipases are well-studied, but few studies have examined the mechanisms of lipase expression regulation. As a global regulatory protein, PmrA controls the expression of multiple genes such as the Dot/Icm apparatus, eukaryotic-like proteins, and secreted effectors. In this study, the effect of PmrA on expression of the lipase lipA in Pseudomonas aeruginosa PAO1 was investigated by knocking out or overexpressing pmrA, rsmY, and rsmA. PmrA regulated the expression of lipA at both the transcriptional and translational level although translation was the pivotal regulatory mechanism for lipA expression. PmrA also regulated the expression of rsmY. Using gel mobility shift assay and pmrA/rsmY double gene knock-out model, we showed that PmrA directly bound to the promoter sequence of rsmY to regulate lipA expression. Translation of lipA was activated by the PmrA/PmrB system via RsmA. Specifically, the Shine-Dalgarno (SD) sequence located at lipA mRNA was overlapped through combination between RsmA and the AGAUGA sequence, subsequently blocking the 30S ribosomal subunit to the SD sequence, leading to translational inhibition of lipA. Transcriptional repression of RsmY initiated translation of lipA through negative translational regulation of rsmA. In conclusion, this study demonstrated that in P. aeruginosa PAO1, PmrA mainly regulated rsmY expression at a translational level to influence lipA expression. RsmY primarily activated lipA translation via negative translational regulation of rsmA.
topic lipase
Pseudomonas aeruginosa
mutation
expression
gene regulation
url http://journal.frontiersin.org/article/10.3389/fmicb.2017.02690/full
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