Association states of the transcription activator protein NtrC from E. coli determined by analytical ultracentrifugation 1 1Edited by K. Yamamoto

Rippe, Karsten; Mücke, Norbert; Schulz, Alexandra

doi:10.1006/jmbi.1998.1746

Cited by 57 publications

(72 citation statements)

References 65 publications

(139 reference statements)

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“…This approach has been used successfully for other protein-protein and protein-DNA interactions (42)(43)(44), although there are notable exceptions (45). Estimated with Equation 5, v P n D is ϳ0.73 for the complexes described here.…”

Section: Methodsmentioning

confidence: 97%

Interactions of Human O6-Alkylguanine-DNA Alkyltransferase (AGT) with Short Single-stranded DNAs

Rasimas

Kar

Pegg

et al. 2007

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 97%

Interactions of Human O6-Alkylguanine-DNA Alkyltransferase (AGT) with Short Single-stranded DNAs

Rasimas

Kar

Pegg

et al. 2007

Journal of Biological Chemistry

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“…rez-Martı! n & de Lorenzo, 1996b), and on the various studies on the archetype of σ&%-dependent enhancer-binding proteins, NtrC (Rippe et al, 1998). Although our former work was done with linear DNA and the work presented here largely involves supercoiled DNA, we did not find any incompatibility in the outcome of the respective experiments.…”

Section: Conclusion : Oligomerization Of Xylr During Activation Of Pumentioning

confidence: 53%

“…6. It is assumed that, similar to NtrC (Rippe et al, 1998), the predominant form of XylR∆A in solution is a dimer. In the absence of ATP, such a dimer has a low affinity for the UAS, with which it interacts non-cooperatively.…”

Section: Conclusion : Oligomerization Of Xylr During Activation Of Pumentioning

confidence: 99%

Visualization of DNA–protein intermediates during activation of the Pu promoter of the TOL plasmid of Pseudomonas putida

Garmendía

2000

Microbiology

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The ATP-dependent multimerization process undergone by the σ 54 -dependent activator XylR of the TOL plasmid pWW0 of Pseudomonas putida when bound to the upstream activating sequences (UAS) of the cognate Pu promoter was examined by transmission electron microscopy (TEM). To this end, supercoiled DNA templates were combined with increasing concentrations of the constitutive XylR variant XylR∆A, with or without ATP or its non-hydrolysable analogue ATPγS, and the resulting complexes were visualized by TEM. The different types of DNA-protein association were analysed and a statistical study of the frequency of the various forms was made. ATP appeared to establish an equilibrium between different molecular associations, as well as major changes in the physical shape of the DNA-protein complexes. The formation of higher nucleoprotein structures frequently bearing DNA bends became manifest. Such complexes often engaged otherwise separated UAScontaining plasmids, indicating that the ATP-driven multimer included XylR molecules recruited in trans. Whilst ATP caused the different types of XylR-DNA complex to occur at quite balanced frequencies, ATPγS appeared to displace the distribution predominantly towards the higher order forms. These data are compatible with the notion that each time ATP is hydrolysed the transcriptional activation complex is disassembled.

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“…The His-tagged proteins listed below were purified by Niaffinity chromatography using the BioLogic LP chromatography system (Bio-Rad). His-s 54 and His-NtrC proteins were overproduced from strains BL21(DE3)(pS54-2) and BL21(DE3)(pNTRC-3), respectively and purified as described by Rippe et al (1997Rippe et al ( , 1998. His-PspF and His-PspFDHTH were obtained by overproduction from SG13009(pREP,pMJ16) and K1527 (pMJ15) respectively and purified according to Jovanovic et al (1999).…”

Section: Methodsmentioning

confidence: 99%

A σ 54-dependent promoter in the regulatory region of the Escherichia coli rpoH gene

et al. 2007

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The Escherichia coli rpoH gene is transcribed from four known and differently regulated promoters: P1, P3, P4 and P5. This study demonstrates that the conserved consensus sequence of the s 54 promoter in the regulatory region of the rpoH gene, described previously, is a functional promoter, P6. The evidence for this conclusion is: (i) the specific binding of the s 54 -RNAP holoenzyme to P6, (ii) the location of the transcription start site at the predicted position (C, 30 nt upstream of ATG) and (iii) the dependence of transcription on s 54 and on an ATP-dependent activator. Nitrogen starvation, heat shock, ethanol and CCCP treatment did not activate transcription from P6 under the conditions examined. Two activators of s 54 promoters, PspF and NtrC, were tested but neither of them acted specifically. Therefore, PspFDHTH, a derivative of PspF, devoid of DNA binding capability but retaining its ATPase activity, was used for transcription in vitro, taking advantage of the relaxed specificity of ATP-dependent activators acting in solution. In experiments in vivo overexpression of PspFDHTH from a plasmid was employed. Thus, the s 54 -dependent transcription capability of the P6 promoter was demonstrated both in vivo and in vitro, although the specific conditions inducing initiation of the transcription remain to be elucidated. The results clearly indicate that the closed s 54 -RNAP-promoter initiation complex was formed in vitro and in vivo and needed only an ATP-dependent activator to start transcription.

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Association states of the transcription activator protein NtrC from E. coli determined by analytical ultracentrifugation 1 1Edited by K. Yamamoto

Cited by 57 publications

References 65 publications

Interactions of Human O6-Alkylguanine-DNA Alkyltransferase (AGT) with Short Single-stranded DNAs

Interactions of Human O6-Alkylguanine-DNA Alkyltransferase (AGT) with Short Single-stranded DNAs

Visualization of DNA–protein intermediates during activation of the Pu promoter of the TOL plasmid of Pseudomonas putida

A σ 54-dependent promoter in the regulatory region of the Escherichia coli rpoH gene

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