Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli

Motility is finely regulated and is crucial to bacterial processes including colonization and biofilm formation. There is a trade-off between motility and growth in bacteria with molecular mechanisms not fully understood. Hypermotile Escherichia coli could be isolated by evolving non-motile cells on...

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Autores principales:	Bingyu Li, Chaofan Hou, Xian Ju, Yong Feng, Zhi-Qiang Ye, Yunzhu Xiao, Mingyao Gu, Chunxiang Fu, Chaoliang Wei, Conghui You
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	evolvable motility spontaneous clpX mutations ClpXP protease protein degradation motility-related reprogramming of gene expression stress response Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/19724f26924b4359b6154108c1dade8c
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spelling	oai:doaj.org-article:19724f26924b4359b6154108c1dade8c2021-11-30T19:03:07ZGain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli2296-418510.3389/fbioe.2021.772397https://doaj.org/article/19724f26924b4359b6154108c1dade8c2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fbioe.2021.772397/fullhttps://doaj.org/toc/2296-4185Motility is finely regulated and is crucial to bacterial processes including colonization and biofilm formation. There is a trade-off between motility and growth in bacteria with molecular mechanisms not fully understood. Hypermotile Escherichia coli could be isolated by evolving non-motile cells on soft agar plates. Most of the isolates carried mutations located upstream of the flhDC promoter region, which upregulate the transcriptional expression of the master regulator of the flagellum biosynthesis, FlhDC. Here, we identified that spontaneous mutations in clpX boosted the motility of E. coli largely, inducing several folds of changes in swimming speed. Among the mutations identified, we further elucidated the molecular mechanism underlying the ClpXV78F mutation on the regulation of E. coli motility. We found that the V78F mutation affected ATP binding to ClpX, resulting in the inability of the mutated ClpXP protease to degrade FlhD as indicated by both structure modeling and in vitro protein degradation assays. Moreover, our proteomic data indicated that the ClpXV78F mutation elevated the stability of known ClpXP targets to various degrees with FlhD as one of the most affected. In addition, the specific tag at the C-terminus of FlhD being recognized for ClpXP degradation was identified. Finally, our transcriptome data characterized that the enhanced expression of the motility genes in the ClpXV78F mutations was intrinsically accompanied by the reduced expression of stress resistance genes relating to the reduced fitness of the hypermotile strains. A similar pattern was observed for previously isolated hypermotile E. coli strains showing high expression of flhDC at the transcriptional level. Hence, clpX appears to be a hot locus comparable to the upstream of the flhDC promoter region evolved to boost bacterial motility, and our finding provides insight into the reduced fitness of the hypermotile bacteria.Bingyu LiBingyu LiBingyu LiChaofan HouXian JuYong FengZhi-Qiang YeYunzhu XiaoMingyao GuChunxiang FuChaoliang WeiConghui YouFrontiers Media S.A.articleevolvable motilityspontaneous clpX mutationsClpXP proteaseprotein degradationmotility-related reprogramming of gene expressionstress responseBiotechnologyTP248.13-248.65ENFrontiers in Bioengineering and Biotechnology, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	evolvable motility spontaneous clpX mutations ClpXP protease protein degradation motility-related reprogramming of gene expression stress response Biotechnology TP248.13-248.65
spellingShingle	evolvable motility spontaneous clpX mutations ClpXP protease protein degradation motility-related reprogramming of gene expression stress response Biotechnology TP248.13-248.65 Bingyu Li Bingyu Li Bingyu Li Chaofan Hou Xian Ju Yong Feng Zhi-Qiang Ye Yunzhu Xiao Mingyao Gu Chunxiang Fu Chaoliang Wei Conghui You Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli
description	Motility is finely regulated and is crucial to bacterial processes including colonization and biofilm formation. There is a trade-off between motility and growth in bacteria with molecular mechanisms not fully understood. Hypermotile Escherichia coli could be isolated by evolving non-motile cells on soft agar plates. Most of the isolates carried mutations located upstream of the flhDC promoter region, which upregulate the transcriptional expression of the master regulator of the flagellum biosynthesis, FlhDC. Here, we identified that spontaneous mutations in clpX boosted the motility of E. coli largely, inducing several folds of changes in swimming speed. Among the mutations identified, we further elucidated the molecular mechanism underlying the ClpXV78F mutation on the regulation of E. coli motility. We found that the V78F mutation affected ATP binding to ClpX, resulting in the inability of the mutated ClpXP protease to degrade FlhD as indicated by both structure modeling and in vitro protein degradation assays. Moreover, our proteomic data indicated that the ClpXV78F mutation elevated the stability of known ClpXP targets to various degrees with FlhD as one of the most affected. In addition, the specific tag at the C-terminus of FlhD being recognized for ClpXP degradation was identified. Finally, our transcriptome data characterized that the enhanced expression of the motility genes in the ClpXV78F mutations was intrinsically accompanied by the reduced expression of stress resistance genes relating to the reduced fitness of the hypermotile strains. A similar pattern was observed for previously isolated hypermotile E. coli strains showing high expression of flhDC at the transcriptional level. Hence, clpX appears to be a hot locus comparable to the upstream of the flhDC promoter region evolved to boost bacterial motility, and our finding provides insight into the reduced fitness of the hypermotile bacteria.
format	article
author	Bingyu Li Bingyu Li Bingyu Li Chaofan Hou Xian Ju Yong Feng Zhi-Qiang Ye Yunzhu Xiao Mingyao Gu Chunxiang Fu Chaoliang Wei Conghui You
author_facet	Bingyu Li Bingyu Li Bingyu Li Chaofan Hou Xian Ju Yong Feng Zhi-Qiang Ye Yunzhu Xiao Mingyao Gu Chunxiang Fu Chaoliang Wei Conghui You
author_sort	Bingyu Li
title	Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli
title_short	Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli
title_full	Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli
title_fullStr	Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli
title_full_unstemmed	Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli
title_sort	gain of spontaneous clpx mutations boosting motility via adaption to environments in escherichia coli
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/19724f26924b4359b6154108c1dade8c
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Gain of Spontaneous clpX Mutations Boosting Motility via Adaption to Environments in Escherichia coli

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