Abstract
The aldA gene (encoding aldehyde dehydrogenase) of Escherichia coli is anaerobically repressed by ArcA-P, the phosphorylated response regulator of the ArcB/A two-component signal transduction system. The promoter region of aldA contains two 10-bp sequences (5'-TGTTAATTAA-3') that perfectly match the proposed ArcA-P binding consensus (5'-[A/T]GTTAATTA[A/T]-3'). One consensus sequence is on the coding strand (-13 to -4 from the transcriptional start point), whereas the other is on the template strand (position -2 to -11). In this study we used the aldA promoter to test the validity of the proposed consensus sequence. DNase I protection experiments confirmed the 10-bp sequence to be a strong ArcA-P binding site. Alteration of the wild-type sequence from 5'-TGTTAATTAAC-3' to 5'-TCTTAATTAAG-3' or 5'-TATTAATTAAT-3' by site-directed mutagenesis markedly decreased the in vitro affinity of the promoter region for ArcA-P, and abolished the anaerobic repression of mutant attλ::Φ (aldA'-lacZ) transcriptional reporter constructs. Both the in vitro and in vivo results therefore support the proposed consensus sequence.
| Original language | English |
|---|---|
| Pages (from-to) | 170-176 |
| Number of pages | 7 |
| Journal | MGG Molecular & General Genetics |
| Volume | 261 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 1999 |
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Keywords
- Aldehyde dehydrogenase
- Arc two-component system
- Escherichia coli
- Signal transduction
ASJC Scopus subject areas
- Genetics
- Molecular Biology
Cite this
A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli. / Pellicer, M. Teresa; Lynch, A. S.; De Wulf, P.; Boyd, D.; Aguilar, Juan; Lin, E. C C.
In: MGG Molecular & General Genetics, Vol. 261, No. 1, 1999, p. 170-176.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli
AU - Pellicer, M. Teresa
AU - Lynch, A. S.
AU - De Wulf, P.
AU - Boyd, D.
AU - Aguilar, Juan
AU - Lin, E. C C
PY - 1999
Y1 - 1999
N2 - The aldA gene (encoding aldehyde dehydrogenase) of Escherichia coli is anaerobically repressed by ArcA-P, the phosphorylated response regulator of the ArcB/A two-component signal transduction system. The promoter region of aldA contains two 10-bp sequences (5'-TGTTAATTAA-3') that perfectly match the proposed ArcA-P binding consensus (5'-[A/T]GTTAATTA[A/T]-3'). One consensus sequence is on the coding strand (-13 to -4 from the transcriptional start point), whereas the other is on the template strand (position -2 to -11). In this study we used the aldA promoter to test the validity of the proposed consensus sequence. DNase I protection experiments confirmed the 10-bp sequence to be a strong ArcA-P binding site. Alteration of the wild-type sequence from 5'-TGTTAATTAAC-3' to 5'-TCTTAATTAAG-3' or 5'-TATTAATTAAT-3' by site-directed mutagenesis markedly decreased the in vitro affinity of the promoter region for ArcA-P, and abolished the anaerobic repression of mutant attλ::Φ (aldA'-lacZ) transcriptional reporter constructs. Both the in vitro and in vivo results therefore support the proposed consensus sequence.
AB - The aldA gene (encoding aldehyde dehydrogenase) of Escherichia coli is anaerobically repressed by ArcA-P, the phosphorylated response regulator of the ArcB/A two-component signal transduction system. The promoter region of aldA contains two 10-bp sequences (5'-TGTTAATTAA-3') that perfectly match the proposed ArcA-P binding consensus (5'-[A/T]GTTAATTA[A/T]-3'). One consensus sequence is on the coding strand (-13 to -4 from the transcriptional start point), whereas the other is on the template strand (position -2 to -11). In this study we used the aldA promoter to test the validity of the proposed consensus sequence. DNase I protection experiments confirmed the 10-bp sequence to be a strong ArcA-P binding site. Alteration of the wild-type sequence from 5'-TGTTAATTAAC-3' to 5'-TCTTAATTAAG-3' or 5'-TATTAATTAAT-3' by site-directed mutagenesis markedly decreased the in vitro affinity of the promoter region for ArcA-P, and abolished the anaerobic repression of mutant attλ::Φ (aldA'-lacZ) transcriptional reporter constructs. Both the in vitro and in vivo results therefore support the proposed consensus sequence.
KW - Aldehyde dehydrogenase
KW - Arc two-component system
KW - Escherichia coli
KW - Signal transduction
UR - http://www.scopus.com/inward/record.url?scp=0032982216&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032982216&partnerID=8YFLogxK
U2 - 10.1007/s004380050954
DO - 10.1007/s004380050954
M3 - Article
VL - 261
SP - 170
EP - 176
JO - Molecular Genetics and Genomics
JF - Molecular Genetics and Genomics
SN - 1617-4615
IS - 1
ER -