Roles of DNA sequence and sigma A factor in transcription of the vraSR operon.

Antoaneta Belcheva, Vidhu Verma, Artyom Korenevsky, Michael Fridman, Krishan Kumar, Dasantila Golemi-Kotra

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Cell wall damage in Staphylococcus aureus induces a rapid genome-wide response, referred to as the cell wall stress stimulon. This response is mediated by a two-component system, the vancomycin resistance-associated sensor/regulator (VraSR). The response regulator protein VraR is a transcription factor. Here, we demonstrate that two VraR binding sites in the vraSR operon control region are involved in the regulation of the vraSR operon. The sites are centered at the -60 and -35 nucleotide positions and are referred to as R1 and R2, respectively. DNase I footprinting and lux operon reporter vector studies showed that both of these sites communicate intimately with each other to fine-tune the activity of the vraSR operon. Mutagenesis of the VraR binding sites showed that dimerization of unphosphorylated VraR at R1 is driven by a hierarchy in VraR binding and by the proximity of the two tandem VraR binding sequences at this site. On the other hand, these studies show that the lack of sequence conservation and the distance between the VraR binding sequences in R2 ensure that VraR is recruited to this site only when phosphorylated (hence, under stress conditions). Furthermore, we demonstrate that sigma A (SigA) factor is involved in the regulation of the vraSR operon. Our study shows that sigma A factor does not bind to the vraSR operon control region in the absence of VraR, suggesting that VraR may interact directly with this factor.
OriginalsprogEngelsk
TidsskriftJournal of Bacteriology
Vol/bind194
Udgave nummer1
Sider (fra-til)61-71
ISSN0021-9193
DOI
StatusUdgivet - 2012
Udgivet eksterntJa

Fingeraftryk

Cell Wall
Vancomycin Resistance
Deoxyribonuclease I
Dimerization
Proteins

Citer dette

Belcheva, A., Verma, V., Korenevsky, A., Fridman, M., Kumar, K., & Golemi-Kotra, D. (2012). Roles of DNA sequence and sigma A factor in transcription of the vraSR operon. Journal of Bacteriology, 194(1), 61-71. https://doi.org/10.1128/JB.06143-11
Belcheva, Antoaneta ; Verma, Vidhu ; Korenevsky, Artyom ; Fridman, Michael ; Kumar, Krishan ; Golemi-Kotra, Dasantila. / Roles of DNA sequence and sigma A factor in transcription of the vraSR operon. I: Journal of Bacteriology. 2012 ; Bind 194, Nr. 1. s. 61-71.
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title = "Roles of DNA sequence and sigma A factor in transcription of the vraSR operon.",
abstract = "Cell wall damage in Staphylococcus aureus induces a rapid genome-wide response, referred to as the cell wall stress stimulon. This response is mediated by a two-component system, the vancomycin resistance-associated sensor/regulator (VraSR). The response regulator protein VraR is a transcription factor. Here, we demonstrate that two VraR binding sites in the vraSR operon control region are involved in the regulation of the vraSR operon. The sites are centered at the -60 and -35 nucleotide positions and are referred to as R1 and R2, respectively. DNase I footprinting and lux operon reporter vector studies showed that both of these sites communicate intimately with each other to fine-tune the activity of the vraSR operon. Mutagenesis of the VraR binding sites showed that dimerization of unphosphorylated VraR at R1 is driven by a hierarchy in VraR binding and by the proximity of the two tandem VraR binding sequences at this site. On the other hand, these studies show that the lack of sequence conservation and the distance between the VraR binding sequences in R2 ensure that VraR is recruited to this site only when phosphorylated (hence, under stress conditions). Furthermore, we demonstrate that sigma A (SigA) factor is involved in the regulation of the vraSR operon. Our study shows that sigma A factor does not bind to the vraSR operon control region in the absence of VraR, suggesting that VraR may interact directly with this factor.",
author = "Antoaneta Belcheva and Vidhu Verma and Artyom Korenevsky and Michael Fridman and Krishan Kumar and Dasantila Golemi-Kotra",
year = "2012",
doi = "10.1128/JB.06143-11",
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pages = "61--71",
journal = "Journal of Bacteriology",
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Belcheva, A, Verma, V, Korenevsky, A, Fridman, M, Kumar, K & Golemi-Kotra, D 2012, 'Roles of DNA sequence and sigma A factor in transcription of the vraSR operon.', Journal of Bacteriology, bind 194, nr. 1, s. 61-71. https://doi.org/10.1128/JB.06143-11

Roles of DNA sequence and sigma A factor in transcription of the vraSR operon. / Belcheva, Antoaneta; Verma, Vidhu; Korenevsky, Artyom; Fridman, Michael; Kumar, Krishan; Golemi-Kotra, Dasantila.

I: Journal of Bacteriology, Bind 194, Nr. 1, 2012, s. 61-71.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Roles of DNA sequence and sigma A factor in transcription of the vraSR operon.

AU - Belcheva, Antoaneta

AU - Verma, Vidhu

AU - Korenevsky, Artyom

AU - Fridman, Michael

AU - Kumar, Krishan

AU - Golemi-Kotra, Dasantila

PY - 2012

Y1 - 2012

N2 - Cell wall damage in Staphylococcus aureus induces a rapid genome-wide response, referred to as the cell wall stress stimulon. This response is mediated by a two-component system, the vancomycin resistance-associated sensor/regulator (VraSR). The response regulator protein VraR is a transcription factor. Here, we demonstrate that two VraR binding sites in the vraSR operon control region are involved in the regulation of the vraSR operon. The sites are centered at the -60 and -35 nucleotide positions and are referred to as R1 and R2, respectively. DNase I footprinting and lux operon reporter vector studies showed that both of these sites communicate intimately with each other to fine-tune the activity of the vraSR operon. Mutagenesis of the VraR binding sites showed that dimerization of unphosphorylated VraR at R1 is driven by a hierarchy in VraR binding and by the proximity of the two tandem VraR binding sequences at this site. On the other hand, these studies show that the lack of sequence conservation and the distance between the VraR binding sequences in R2 ensure that VraR is recruited to this site only when phosphorylated (hence, under stress conditions). Furthermore, we demonstrate that sigma A (SigA) factor is involved in the regulation of the vraSR operon. Our study shows that sigma A factor does not bind to the vraSR operon control region in the absence of VraR, suggesting that VraR may interact directly with this factor.

AB - Cell wall damage in Staphylococcus aureus induces a rapid genome-wide response, referred to as the cell wall stress stimulon. This response is mediated by a two-component system, the vancomycin resistance-associated sensor/regulator (VraSR). The response regulator protein VraR is a transcription factor. Here, we demonstrate that two VraR binding sites in the vraSR operon control region are involved in the regulation of the vraSR operon. The sites are centered at the -60 and -35 nucleotide positions and are referred to as R1 and R2, respectively. DNase I footprinting and lux operon reporter vector studies showed that both of these sites communicate intimately with each other to fine-tune the activity of the vraSR operon. Mutagenesis of the VraR binding sites showed that dimerization of unphosphorylated VraR at R1 is driven by a hierarchy in VraR binding and by the proximity of the two tandem VraR binding sequences at this site. On the other hand, these studies show that the lack of sequence conservation and the distance between the VraR binding sequences in R2 ensure that VraR is recruited to this site only when phosphorylated (hence, under stress conditions). Furthermore, we demonstrate that sigma A (SigA) factor is involved in the regulation of the vraSR operon. Our study shows that sigma A factor does not bind to the vraSR operon control region in the absence of VraR, suggesting that VraR may interact directly with this factor.

U2 - 10.1128/JB.06143-11

DO - 10.1128/JB.06143-11

M3 - Journal article

VL - 194

SP - 61

EP - 71

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 1

ER -

Belcheva A, Verma V, Korenevsky A, Fridman M, Kumar K, Golemi-Kotra D. Roles of DNA sequence and sigma A factor in transcription of the vraSR operon. Journal of Bacteriology. 2012;194(1):61-71. https://doi.org/10.1128/JB.06143-11