Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400

Nguyen, Tinh T.; Martí-Arbona, Ricardo; Hall, Richard S.; Maity, Tuhin; Valdez, Yolanda E.; Dunbar, John M.; Unkefer, Clifford J.; Unkefer, Pat J.

doi:10.5539/jmbr.v3n1p37

Title: Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400

Abstract

Transcriptional regulators (TRs) are an important and versatile group of proteins, yet very little progress has been achieved towards the discovery and annotation of their biological functions. We have characterized a previously unknown organic hydroperoxide resistance regulator from Burkholderia xenovoransLB400, Bxe_B2842, which is homologous to E. coli’s OhrR. Bxe_B2842 regulates the expression of an organic hydroperoxide resistance protein (OsmC). We utilized frontal affinity chromatography coupled with mass spectrometry (FAC-MS) and electrophoretic mobility gel shift assays (EMSA) to identify and characterize the possible effectors of the regulation by Bxe_B2842. Without an effector, Bxe_B2842 binds a DNA operator sequence (DOS) upstream of osmC. FAC-MS results suggest that 2-aminophenol binds to the protein and is potentially an effector molecule. EMSA analysis shows that 2-aminophenol also attenuates the Bxe_B2842’s affinity for its DOS. EMSA analysis also shows that organic peroxides attenuate Bxe_B2842/DOS affinity, suggesting that binding of the TR to its DOS is regulated by the two-cysteine mechanism, common to TRs in this family. Bxe_B2842 is the first OhrR TR to have both oxidative and effector-binding mechanisms of regulation. Our paper reveals further mechanistic diversity TR mediated gene regulation and provides insights into methods for function discovery of TRs.

Authors:

Nguyen, Tinh T. ^[1]; Martí-Arbona, Ricardo ^[1]; Hall, Richard S. ^[1]; Maity, Tuhin ^[1]; Valdez, Yolanda E. ^[2]; Dunbar, John M. ^[1]; Unkefer, Clifford J. ^[1]; Unkefer, Pat J. ^[1]

Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)ry

Publication Date:: Tue May 21 00:00:00 EDT 2013

Research Org.:: Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)

Sponsoring Org.:: USDOE

OSTI Identifier:: 1224042

Report Number(s):: LA-UR-13-21680
Journal ID: ISSN 1925-430X

Grant/Contract Number:: AC52-06NA25396

Resource Type:: Accepted Manuscript

Journal Name:: Journal of Molecular Biology Research

Additional Journal Information:: Journal Volume: 3; Journal Issue: 1; Journal ID: ISSN 1925-430X

Publisher:: Canadian Center of Science and Education

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; transcriptional regulator (TR), transcriptional regulator effector (TRE); fontal affinity chromatography coupled to mass spectrometry (FAC-MS), Burkholderia xenovorans LB400; organic hydroperoxide resistance regulator (OhrR); derepression; organic peroxides; Bxe_B2842 and 2-aminophenol

Citation Formats


                    Nguyen, Tinh T., Martí-Arbona, Ricardo, Hall, Richard S., Maity, Tuhin, Valdez, Yolanda E., Dunbar, John M., Unkefer, Clifford J., and Unkefer, Pat J. Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400.  United States: N. p., 2013. 
Web.  doi:10.5539/jmbr.v3n1p37.

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                    Nguyen, Tinh T., Martí-Arbona, Ricardo, Hall, Richard S., Maity, Tuhin, Valdez, Yolanda E., Dunbar, John M., Unkefer, Clifford J., & Unkefer, Pat J. Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400.  United States.  https://doi.org/10.5539/jmbr.v3n1p37

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                    Nguyen, Tinh T., Martí-Arbona, Ricardo, Hall, Richard S., Maity, Tuhin, Valdez, Yolanda E., Dunbar, John M., Unkefer, Clifford J., and Unkefer, Pat J. Tue .  
"Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400".  United States.  https://doi.org/10.5539/jmbr.v3n1p37.  https://www.osti.gov/servlets/purl/1224042.

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@article{osti_1224042,

  title        = {Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400},

  author       = {Nguyen, Tinh T. and Martí-Arbona, Ricardo and Hall, Richard S. and Maity, Tuhin and Valdez, Yolanda E. and Dunbar, John M. and Unkefer, Clifford J. and Unkefer, Pat J.},

  abstractNote = {Transcriptional regulators (TRs) are an important and versatile group of proteins, yet very little progress has been achieved towards the discovery and annotation of their biological functions. We have characterized a previously unknown organic hydroperoxide resistance regulator from Burkholderia xenovoransLB400, Bxe_B2842, which is homologous to E. coli’s OhrR. Bxe_B2842 regulates the expression of an organic hydroperoxide resistance protein (OsmC). We utilized frontal affinity chromatography coupled with mass spectrometry (FAC-MS) and electrophoretic mobility gel shift assays (EMSA) to identify and characterize the possible effectors of the regulation by Bxe_B2842. Without an effector, Bxe_B2842 binds a DNA operator sequence (DOS) upstream of osmC. FAC-MS results suggest that 2-aminophenol binds to the protein and is potentially an effector molecule. EMSA analysis shows that 2-aminophenol also attenuates the Bxe_B2842’s affinity for its DOS. EMSA analysis also shows that organic peroxides attenuate Bxe_B2842/DOS affinity, suggesting that binding of the TR to its DOS is regulated by the two-cysteine mechanism, common to TRs in this family. Bxe_B2842 is the first OhrR TR to have both oxidative and effector-binding mechanisms of regulation. Our paper reveals further mechanistic diversity TR mediated gene regulation and provides insights into methods for function discovery of TRs.},

  doi          = {10.5539/jmbr.v3n1p37},

  journal      = {Journal of Molecular Biology Research},

  number       = 1,

  volume       = 3,

  place        = {United States},

  year         = {Tue May 21 00:00:00 EDT 2013},

  month        = {Tue May 21 00:00:00 EDT 2013}

}

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Title: Identification and In-vivo Characterization of a Novel OhrR Transcriptional Regulator in Burkholderia xenovorans LB400

Abstract

Citation Formats

Automated genomic context analysis and experimental validation platform for discovery of prokaryote transcriptional regulator functions journal, January 2014

Automated genomic context analysis and experimental validation platform for discovery of prokaryote transcriptional regulator functions
journal, January 2014