Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator

Abstract The transmembrane DNA-binding protein CadC of E. coli, a representative of the ToxR-like receptor family, combines input and effector domains for signal sensing and transcriptional activation, respectively, in a single protein, thus representing one of the simplest signalling systems. At ac...

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Autores principales:	Andreas Schlundt, Sophie Buchner, Robert Janowski, Thomas Heydenreich, Ralf Heermann, Jürgen Lassak, Arie Geerlof, Ralf Stehle, Dierk Niessing, Kirsten Jung, Michael Sattler
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a3f5cd158ad24f1faf99b7832603d2e3
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spelling	oai:doaj.org-article:a3f5cd158ad24f1faf99b7832603d2e32021-12-02T16:08:19ZStructure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator10.1038/s41598-017-01031-92045-2322https://doaj.org/article/a3f5cd158ad24f1faf99b7832603d2e32017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01031-9https://doaj.org/toc/2045-2322Abstract The transmembrane DNA-binding protein CadC of E. coli, a representative of the ToxR-like receptor family, combines input and effector domains for signal sensing and transcriptional activation, respectively, in a single protein, thus representing one of the simplest signalling systems. At acidic pH in a lysine-rich environment, CadC activates the transcription of the cadBA operon through recruitment of the RNA polymerase (RNAP) to the two cadBA promoter sites, Cad1 and Cad2, which are directly bound by CadC. However, the molecular details for its interaction with DNA have remained elusive. Here, we present the crystal structure of the CadC DNA-binding domain (DBD) and show that it adopts a winged helix-turn-helix fold. The interaction with the cadBA promoter site Cad1 is studied by using nuclear magnetic resonance (NMR) spectroscopy, biophysical methods and functional assays and reveals a preference for AT-rich regions. By mutational analysis we identify amino acids within the CadC DBD that are crucial for DNA-binding and functional activity. Experimentally derived structural models of the CadC-DNA complex indicate that the CadC DBD employs mainly non-sequence-specific over a few specific contacts. Our data provide molecular insights into the CadC-DNA interaction and suggest how CadC dimerization may provide high-affinity binding to the Cad1 promoter.Andreas SchlundtSophie BuchnerRobert JanowskiThomas HeydenreichRalf HeermannJürgen LassakArie GeerlofRalf StehleDierk NiessingKirsten JungMichael SattlerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-16 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Andreas Schlundt Sophie Buchner Robert Janowski Thomas Heydenreich Ralf Heermann Jürgen Lassak Arie Geerlof Ralf Stehle Dierk Niessing Kirsten Jung Michael Sattler Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator
description	Abstract The transmembrane DNA-binding protein CadC of E. coli, a representative of the ToxR-like receptor family, combines input and effector domains for signal sensing and transcriptional activation, respectively, in a single protein, thus representing one of the simplest signalling systems. At acidic pH in a lysine-rich environment, CadC activates the transcription of the cadBA operon through recruitment of the RNA polymerase (RNAP) to the two cadBA promoter sites, Cad1 and Cad2, which are directly bound by CadC. However, the molecular details for its interaction with DNA have remained elusive. Here, we present the crystal structure of the CadC DNA-binding domain (DBD) and show that it adopts a winged helix-turn-helix fold. The interaction with the cadBA promoter site Cad1 is studied by using nuclear magnetic resonance (NMR) spectroscopy, biophysical methods and functional assays and reveals a preference for AT-rich regions. By mutational analysis we identify amino acids within the CadC DBD that are crucial for DNA-binding and functional activity. Experimentally derived structural models of the CadC-DNA complex indicate that the CadC DBD employs mainly non-sequence-specific over a few specific contacts. Our data provide molecular insights into the CadC-DNA interaction and suggest how CadC dimerization may provide high-affinity binding to the Cad1 promoter.
format	article
author	Andreas Schlundt Sophie Buchner Robert Janowski Thomas Heydenreich Ralf Heermann Jürgen Lassak Arie Geerlof Ralf Stehle Dierk Niessing Kirsten Jung Michael Sattler
author_facet	Andreas Schlundt Sophie Buchner Robert Janowski Thomas Heydenreich Ralf Heermann Jürgen Lassak Arie Geerlof Ralf Stehle Dierk Niessing Kirsten Jung Michael Sattler
author_sort	Andreas Schlundt
title	Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator
title_short	Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator
title_full	Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator
title_fullStr	Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator
title_full_unstemmed	Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator
title_sort	structure-function analysis of the dna-binding domain of a transmembrane transcriptional activator
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/a3f5cd158ad24f1faf99b7832603d2e3
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Structure-function analysis of the DNA-binding domain of a transmembrane transcriptional activator

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