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eLife Volume 8 ,2019-12-03
Crl activates transcription by stabilizing active conformation of the master stress transcription initiation factor
Chromosomes and Gene Expression
Juncao Xu 1 , 2 Kaijie Cui 2 , 3 Liqiang Shen 1 , 2 Jing Shi 4 , 5 Lingting Li 1 , 2 Linlin You 1 , 2 Chengli Fang 1 , 2 Guoping Zhao 1 , 6 , 7 , 8 , 9 Yu Feng 4 , 5 Bei Yang 3 Yu Zhang 1
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DOI:10.7554/eLife.50928
Received 2019-08-07, accepted for publication 2019-12-03, Published 2019-12-03
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摘要

σS is a master transcription initiation factor that protects bacterial cells from various harmful environmental stresses including antibiotic pressure. Although its mechanism remains unclear, it is known that full activation of σS-mediated transcription requires a σS-specific activator, Crl. In this study, we determined a 3.80 Å cryo-EM structure of an Escherichia coli transcription activation complex (E. coli Crl-TAC) comprising E. coli σS-RNA polymerase (σS-RNAP) holoenzyme, Crl, and a nucleic-acid scaffold. The structure reveals that Crl interacts with domain 2 of σS (σS2) and the RNAP core enzyme, but does not contact promoter DNA. Results from subsequent hydrogen-deuterium exchange mass spectrometry (HDX-MS) indicate that Crl stabilizes key structural motifs within σS2 to promote the assembly of the σS-RNAP holoenzyme and also to facilitate formation of an RNA polymerase–promoter DNA open complex (RPo). Our study demonstrates a unique DNA contact-independent mechanism of transcription activation, thereby defining a previously unrecognized mode of transcription activation in cells.

关键词

E. coli;stress response;sigma factor;transcription activation;Crl;RNA polymerase;transcription factors

授权许可

© 2019, Xu et al
http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Juncao Xu,Kaijie Cui,Liqiang Shen,Jing Shi,Lingting Li,Linlin You,Chengli Fang,Guoping Zhao,Yu Feng,Bei Yang,Yu Zhang. Crl activates transcription by stabilizing active conformation of the master stress transcription initiation factor. eLife ,Vol.8(2019)

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