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.

A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli / Pellicer, M. T.; Lynch, A. S.; De Wulf, P.; Boyd, D.; Aguilar, J.; Lin, E. C. C.. - In: MOLECULAR AND GENERAL GENETICS. - ISSN 0026-8925. - STAMPA. - 1999, vol. 261:1(1999), pp. 170-176. [10.1007/s004380050954]

A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli

De Wulf P.;
1999-01-01

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.
1999
1
Pellicer, M. T.; Lynch, A. S.; De Wulf, P.; Boyd, D.; Aguilar, J.; Lin, E. C. C.
A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli / Pellicer, M. T.; Lynch, A. S.; De Wulf, P.; Boyd, D.; Aguilar, J.; Lin, E. C. C.. - In: MOLECULAR AND GENERAL GENETICS. - ISSN 0026-8925. - STAMPA. - 1999, vol. 261:1(1999), pp. 170-176. [10.1007/s004380050954]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11572/262009
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