PMID:10694378
| Citation |
Rippe, K (2000) Simultaneous binding of two DNA duplexes to the NtrC-enhancer complex studied by two-color fluorescence cross-correlation spectroscopy. Biochemistry 39:2131-9 |
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| Abstract |
The transcription activator protein NtrC (nitrogen regulatory protein C, also termed NR(I)) can catalyze the transition of Escherichia coli RNA polymerase complexed with the sigma(54) factor (RNAP x sigma(54)) from the closed complex (RNAP x sigma(54) bound at the promoter) to the open complex (melting of the promoter DNA). This process involves phosphorylation of NtrC (NtrC-P), assembly of an octameric NtrC-P complex at the enhancer DNA sequence, interaction of this complex with promoter-bound RNAP x sigma(54) via DNA looping, and hydrolysis of ATP. Here it is demonstrated by two-color fluorescence cross-correlation spectroscopy measurements of 6-carboxyfluorescein and 6-carboxy-X-rhodamine-labeled DNA oligonucleotide duplexes that the NtrC-P complex can bind two DNA duplexes simultaneously. This suggests a model for the conformation of the looped intermediate that is formed between NtrC-P and RNAP. sigma(54) at the glnAp2 promoter during the activation process. |
| Links | |
| Keywords |
Bacterial Proteins/chemistry; Bacterial Proteins/metabolism; Base Sequence; DNA, Bacterial/chemistry; DNA, Bacterial/metabolism; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/metabolism; Diffusion; Enhancer Elements, Genetic; Escherichia coli/chemistry; Escherichia coli/genetics; Escherichia coli Proteins; Models, Chemical; Models, Molecular; Molecular Sequence Data; Nucleic Acid Heteroduplexes/chemistry; Nucleic Acid Heteroduplexes/metabolism; PII Nitrogen Regulatory Proteins; Phosphorylation; Protein Binding; Reproducibility of Results; Spectrometry, Fluorescence/methods; Trans-Activators; Transcription Factors |
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