Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1

Transition metals, such as cadmium (Cd) and zinc (Zn), can be detrimental to cell viability in excess. Bacteria contain conserved regulatory mechanisms to sense and respond to a variety of environmental stressors, such as an influx of metal cations. One such mechanism is the activation of metalloreg...

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Main Authors: John K. Barrows, Ross M. Wood, Alaina B. Westee, Kamya A. Stubbs, Melanie Ratliff-Griffin, Michael W. Van Dyke
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1608957/full
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author John K. Barrows
Ross M. Wood
Alaina B. Westee
Kamya A. Stubbs
Melanie Ratliff-Griffin
Michael W. Van Dyke
author_facet John K. Barrows
Ross M. Wood
Alaina B. Westee
Kamya A. Stubbs
Melanie Ratliff-Griffin
Michael W. Van Dyke
author_sort John K. Barrows
collection DOAJ
description Transition metals, such as cadmium (Cd) and zinc (Zn), can be detrimental to cell viability in excess. Bacteria contain conserved regulatory mechanisms to sense and respond to a variety of environmental stressors, such as an influx of metal cations. One such mechanism is the activation of metalloregulatory transcription factors that bind to cognate metal cofactors to induce a transcription regulatory response. Metalloregulatory transcription factor families, such as the ferric uptake regulator (FUR), mercury-resistant regulator (MerR), copper-sensitive operon repressor (CsoR), and diphtheria toxin regulator (DtxR), are found widespread throughout bacterial genomes. Often, these transcription factors bind a specific DNA sequence found in the promoter of regulated genes to exert their transcription regulatory functions. In this study, we use an iterative selection technique called restriction endonuclease protection, selection, and amplification (RESPA) to identify the preferred DNA binding sequence for the MerR family, Cd/Zn-responsive regulator, CadR, from the opportunistic human pathogen, Pseudomonas aeruginosa. By doing so, we identify the transcription regulatory network for CadR, which includes the Cd/Zn-exporter, cadA, as well as an uncharacterized zinc ribbon domain-containing protein.
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spelling doaj-art-20450288129646dea10e5ef8a10092852025-08-20T03:49:56ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.16089571608957Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1John K. Barrows0Ross M. Wood1Alaina B. Westee2Kamya A. Stubbs3Melanie Ratliff-Griffin4Michael W. Van Dyke5College of Humanities, Sciences, and Technology, Reinhardt University, Waleska, GA, United StatesDepartment of Cellular and Molecular Biology, Kennesaw State University, Kennesaw, GA, United StatesDepartment of Chemistry and Biochemistry, Kennesaw State University, Kennesaw, GA, United StatesDepartment of Chemistry and Biochemistry, Kennesaw State University, Kennesaw, GA, United StatesDepartment of Cellular and Molecular Biology, Kennesaw State University, Kennesaw, GA, United StatesDepartment of Chemistry and Biochemistry, Kennesaw State University, Kennesaw, GA, United StatesTransition metals, such as cadmium (Cd) and zinc (Zn), can be detrimental to cell viability in excess. Bacteria contain conserved regulatory mechanisms to sense and respond to a variety of environmental stressors, such as an influx of metal cations. One such mechanism is the activation of metalloregulatory transcription factors that bind to cognate metal cofactors to induce a transcription regulatory response. Metalloregulatory transcription factor families, such as the ferric uptake regulator (FUR), mercury-resistant regulator (MerR), copper-sensitive operon repressor (CsoR), and diphtheria toxin regulator (DtxR), are found widespread throughout bacterial genomes. Often, these transcription factors bind a specific DNA sequence found in the promoter of regulated genes to exert their transcription regulatory functions. In this study, we use an iterative selection technique called restriction endonuclease protection, selection, and amplification (RESPA) to identify the preferred DNA binding sequence for the MerR family, Cd/Zn-responsive regulator, CadR, from the opportunistic human pathogen, Pseudomonas aeruginosa. By doing so, we identify the transcription regulatory network for CadR, which includes the Cd/Zn-exporter, cadA, as well as an uncharacterized zinc ribbon domain-containing protein.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1608957/fullgene regulationCadRtranscription factorsmetal-sensingPseudomonas aeruginosa
spellingShingle John K. Barrows
Ross M. Wood
Alaina B. Westee
Kamya A. Stubbs
Melanie Ratliff-Griffin
Michael W. Van Dyke
Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1
Frontiers in Microbiology
gene regulation
CadR
transcription factors
metal-sensing
Pseudomonas aeruginosa
title Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1
title_full Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1
title_fullStr Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1
title_full_unstemmed Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1
title_short Identification of a preferred DNA binding sequence and novel regulon member for CadR in Pseudomonas aeruginosa PAO1
title_sort identification of a preferred dna binding sequence and novel regulon member for cadr in pseudomonas aeruginosa pao1
topic gene regulation
CadR
transcription factors
metal-sensing
Pseudomonas aeruginosa
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1608957/full
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