CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.

The expression of σ(54)-dependent Pseudomonas putida Pu promoter is activated by XylR activator when cells are exposed to a variety of aromatic inducers. In this study, the transcriptional activation of the P. putida Pu promoter was recreated in the heterologous host Escherichia coli. Here we show t...

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Autores principales:	Yuan-Tao Zhang, Feng Jiang, Zhe-Xian Tian, Yi-Xin Huo, Yi-Cheng Sun, Yi-Ping Wang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c7e0c55b0b0a470c9e34e8150b528529
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spelling	oai:doaj.org-article:c7e0c55b0b0a470c9e34e8150b5285292021-11-18T08:36:04ZCRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.1932-620310.1371/journal.pone.0086727https://doaj.org/article/c7e0c55b0b0a470c9e34e8150b5285292014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24466213/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The expression of σ(54)-dependent Pseudomonas putida Pu promoter is activated by XylR activator when cells are exposed to a variety of aromatic inducers. In this study, the transcriptional activation of the P. putida Pu promoter was recreated in the heterologous host Escherichia coli. Here we show that the cAMP receptor protein (CRP), a well-known carbon utilization regulator, had an inhibitory effect on the expression of Pu promoter in a cAMP-dependent manner. The inhibitory effect was not activator specific. In vivo KMnO4 and DMS footprinting analysis indicated that CRP-cAMP poised the RNA polymerase at Pu promoter, inhibiting the isomerization step of the transcription initiation even in the presence of an activator. Therefore, the presence of PTS-sugar, which eliminates cAMP, could activate the poised RNA polymerase at Pu promoter to transcribe. Moreover, the activation region 1 (AR1) of CRP, which interacts directly with the αCTD (C-terminal domain of α-subunit) of RNA polymerase, was found essential for the CRP-mediated inhibition at Pu promoter. A model for the above observations is discussed.Yuan-Tao ZhangFeng JiangZhe-Xian TianYi-Xin HuoYi-Cheng SunYi-Ping WangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 1, p e86727 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yuan-Tao Zhang Feng Jiang Zhe-Xian Tian Yi-Xin Huo Yi-Cheng Sun Yi-Ping Wang CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.
description	The expression of σ(54)-dependent Pseudomonas putida Pu promoter is activated by XylR activator when cells are exposed to a variety of aromatic inducers. In this study, the transcriptional activation of the P. putida Pu promoter was recreated in the heterologous host Escherichia coli. Here we show that the cAMP receptor protein (CRP), a well-known carbon utilization regulator, had an inhibitory effect on the expression of Pu promoter in a cAMP-dependent manner. The inhibitory effect was not activator specific. In vivo KMnO4 and DMS footprinting analysis indicated that CRP-cAMP poised the RNA polymerase at Pu promoter, inhibiting the isomerization step of the transcription initiation even in the presence of an activator. Therefore, the presence of PTS-sugar, which eliminates cAMP, could activate the poised RNA polymerase at Pu promoter to transcribe. Moreover, the activation region 1 (AR1) of CRP, which interacts directly with the αCTD (C-terminal domain of α-subunit) of RNA polymerase, was found essential for the CRP-mediated inhibition at Pu promoter. A model for the above observations is discussed.
format	article
author	Yuan-Tao Zhang Feng Jiang Zhe-Xian Tian Yi-Xin Huo Yi-Cheng Sun Yi-Ping Wang
author_facet	Yuan-Tao Zhang Feng Jiang Zhe-Xian Tian Yi-Xin Huo Yi-Cheng Sun Yi-Ping Wang
author_sort	Yuan-Tao Zhang
title	CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.
title_short	CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.
title_full	CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.
title_fullStr	CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.
title_full_unstemmed	CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.
title_sort	crp-cyclic amp dependent inhibition of the xylene-responsive σ(54)-promoter pu in escherichia coli.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/c7e0c55b0b0a470c9e34e8150b528529
work_keys_str_mv	AT yuantaozhang crpcyclicampdependentinhibitionofthexyleneresponsives54promoterpuinescherichiacoli AT fengjiang crpcyclicampdependentinhibitionofthexyleneresponsives54promoterpuinescherichiacoli AT zhexiantian crpcyclicampdependentinhibitionofthexyleneresponsives54promoterpuinescherichiacoli AT yixinhuo crpcyclicampdependentinhibitionofthexyleneresponsives54promoterpuinescherichiacoli AT yichengsun crpcyclicampdependentinhibitionofthexyleneresponsives54promoterpuinescherichiacoli AT yipingwang crpcyclicampdependentinhibitionofthexyleneresponsives54promoterpuinescherichiacoli
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CRP-cyclic AMP dependent inhibition of the xylene-responsive σ(54)-promoter Pu in Escherichia coli.

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