2uye

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{{STRUCTURE_2uye| PDB=2uye | SCENE= }}
{{STRUCTURE_2uye| PDB=2uye | SCENE= }}
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'''DOUBLE MUTANT Y110S,F111V DNTR FROM BURKHOLDERIA SP. STRAIN DNT IN COMPLEX WITH THIOCYANATE'''
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===DOUBLE MUTANT Y110S,F111V DNTR FROM BURKHOLDERIA SP. STRAIN DNT IN COMPLEX WITH THIOCYANATE===
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==Overview==
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DntR is a bacterial transcription factor that has been isolated from Burkholderia species that are able to degrade the nitro-aromatic compound 2,4-dinitrotoluene. We recently solved the X-ray crystal structure of DntR, which suggested a putative location of an inducer-binding cavity (IBC). In this study, we constructed mutants of DntR in which residues lining the proposed IBC were modified in order to identify the structural elements involved in inducer binding, to modulate the inducer binding specificity, and to investigate the mechanism of transcriptional regulation by DntR. The transcriptional activation of the reporter gene gfp induced by the wild-type and mutant DntRs was monitored by analysing whole-cell fluorescence using flow-cytometry after addition of a number of potential inducer compounds. Three of the mutant proteins (F111L; F111V/H169V and Y110S/F111V) were purified and the binding constants for several of the potential inducers to these mutants were estimated. Furthermore, crystal structures of the F111L and Y110S/F111V mutant proteins were solved and used to explain changes in the inducer binding specificity at an atomic level. A comparison of the inducing capability in the whole-cell system and binding constants for a number of potential inducers suggests a mechanism where binding of an inducer molecule is not the sole requirement for transcriptional activation. In addition, specific interactions between DntR and the inducer molecule resulting in a conformational change of the protein are needed.
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==About this Structure==
==About this Structure==
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[[Category: Transcription regulation]]
[[Category: Transcription regulation]]
[[Category: Transcriptional regulator]]
[[Category: Transcriptional regulator]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Jul 27 19:31:33 2008''

Revision as of 16:31, 27 July 2008

Template:STRUCTURE 2uye

DOUBLE MUTANT Y110S,F111V DNTR FROM BURKHOLDERIA SP. STRAIN DNT IN COMPLEX WITH THIOCYANATE

Template:ABSTRACT PUBMED 17681542

About this Structure

2UYE is a Single protein structure of sequence from Burkholderia cepacia. Full crystallographic information is available from OCA.

Reference

In vivo and in vitro investigation of transcriptional regulation by DntR., Lonneborg R, Smirnova I, Dian C, Leonard GA, Brzezinski P, J Mol Biol. 2007 Sep 21;372(3):571-82. Epub 2007 Jul 3. PMID:17681542

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