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| - | [[Image:2b57.gif|left|200px]] | + | {{Seed}} |
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| | {{STRUCTURE_2b57| PDB=2b57 | SCENE= }} | | {{STRUCTURE_2b57| PDB=2b57 | SCENE= }} |
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| - | '''Guanine Riboswitch C74U mutant bound to 2,6-diaminopurine'''
| + | ===Guanine Riboswitch C74U mutant bound to 2,6-diaminopurine=== |
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| - | ==Overview==
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| - | Riboswitches are cis-acting genetic regulatory elements found commonly in bacterial mRNAs that consist of a metabolite-responsive aptamer domain coupled to a regulatory switch. Purine riboswitches respond to intracellular concentrations of either adenine or guanine/hypoxanthine to control gene expression. The aptamer domain of the purine riboswitch contains a pyrimidine residue (Y74) that forms a Watson-Crick base-pairing interaction with the bound purine nucleobase ligand that discriminates between adenine and guanine. We sought to understand the structural basis of this specificity and the mechanism of ligand recognition by the purine riboswitch. Here, we present the 2,6-diaminopurine-bound structure of a C74U mutant of the xpt-pbuX guanine riboswitch, along with a detailed thermodynamic and kinetic analysis of nucleobase recognition by both the native and mutant riboswitches. These studies demonstrate clearly that the pyrimidine at position 74 is the sole determinant of purine riboswitch specificity. In addition, the mutant riboswitch binds adenine and adenine derivatives well compared with the guanine-responsive riboswitch. Under our experimental conditions, 2,6-diaminopurine binds the RNA with DeltaH=-40.3 kcal mol(-1), DeltaS=-97.6 cal mol(-1)K(-1), and DeltaG=-10.73 kcal mol(-1). A kinetic determination of the slow rate (0.15 x 10(5)M(-1)s(-1) and 2.1 x 10(5)mM(-1)s(-1) for 2-aminopurine binding the adenine-responsive mutant riboswitch and 7-deazaguanine-binding guanine riboswitch, respectively) of association under varying experimental conditions allowed us to propose a mechanism for ligand recognition by the purine riboswitch. A conformationally dynamic unliganded state for the binding pocket is stabilized first by the Watson-Crick base pairing between the ligand and Y74, and by the subsequent ordering of the J2/3 loop, enclosing the ligand within the three-way junction.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_16650860}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 16650860 is the PubMed ID number. |
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| | + | {{ABSTRACT_PUBMED_16650860}} |
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| | ==About this Structure== | | ==About this Structure== |
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| | [[Category: Purine]] | | [[Category: Purine]] |
| | [[Category: Rna-ligand complex]] | | [[Category: Rna-ligand complex]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 19:52:12 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 16:30:53 2008'' |
Revision as of 13:30, 28 July 2008
Template:STRUCTURE 2b57
Guanine Riboswitch C74U mutant bound to 2,6-diaminopurine
Template:ABSTRACT PUBMED 16650860
About this Structure
2B57 is a Single protein structure. Full crystallographic information is available from OCA.
Reference
Thermodynamic and kinetic characterization of ligand binding to the purine riboswitch aptamer domain., Gilbert SD, Stoddard CD, Wise SJ, Batey RT, J Mol Biol. 2006 Jun 9;359(3):754-68. Epub 2006 Apr 21. PMID:16650860
Page seeded by OCA on Mon Jul 28 16:30:53 2008
