1rmn
From Proteopedia
(New page: 200px<br /><applet load="1rmn" size="450" color="white" frame="true" align="right" spinBox="true" caption="1rmn" /> '''A THREE-DIMENSIONAL MODEL FOR THE HAMMERHEAD...) |
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| - | [[Image:1rmn.gif|left|200px]]<br /><applet load="1rmn" size=" | + | [[Image:1rmn.gif|left|200px]]<br /><applet load="1rmn" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1rmn" /> | caption="1rmn" /> | ||
'''A THREE-DIMENSIONAL MODEL FOR THE HAMMERHEAD RIBOZYME BASED ON FLUORESCENCE MEASUREMENTS'''<br /> | '''A THREE-DIMENSIONAL MODEL FOR THE HAMMERHEAD RIBOZYME BASED ON FLUORESCENCE MEASUREMENTS'''<br /> | ||
==Overview== | ==Overview== | ||
| - | For the understanding of the catalytic function of the RNA hammerhead | + | For the understanding of the catalytic function of the RNA hammerhead ribozyme, a three-dimensional model is essential but neither a crystal nor a solution structure has been available. Fluorescence resonance energy transfer (FRET) was used to study the structure of the ribozyme in solution in order to establish the relative spatial orientation of the three constituent Watson-Crick base-paired helical segments. Synthetic constructs were labeled with the fluorescence donor (5-carboxyfluorescein) and acceptor (5-carboxytetramethylrhodamine) located at the ends of the strands constituting the ribozyme molecule. The acceptor helix in helix pairs I and III and in II and III was varied in length from 5 to 11 and 5 to 9 base pairs, respectively, and the FRET efficiencies were determined and correlated with a reference set of labeled RNA duplexes. The FRET efficiencies were predicted on the basis of vector algebra analysis, as a function of the relative helical orientations in the ribozyme constructs, and compared with experimental values. The data were consistent with a Y-shaped arrangement of the ribozyme with helices I and II in close proximity and helix III pointing away. These orientational constraints were used for molecular modeling of a three-dimensional structure of the complete ribozyme. |
==About this Structure== | ==About this Structure== | ||
| - | 1RMN is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/ ]. Full crystallographic information is available from [http:// | + | 1RMN is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/ ]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1RMN OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Eckstein, F.]] | [[Category: Eckstein, F.]] | ||
[[Category: Gohlke, C.]] | [[Category: Gohlke, C.]] | ||
| - | [[Category: Jovin, T | + | [[Category: Jovin, T M.]] |
[[Category: Tuschl, T.]] | [[Category: Tuschl, T.]] | ||
[[Category: Westhof, E.]] | [[Category: Westhof, E.]] | ||
[[Category: ribonucleic acid(hammerhead ribozyme)]] | [[Category: ribonucleic acid(hammerhead ribozyme)]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:52:32 2008'' |
Revision as of 12:52, 21 February 2008
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A THREE-DIMENSIONAL MODEL FOR THE HAMMERHEAD RIBOZYME BASED ON FLUORESCENCE MEASUREMENTS
Overview
For the understanding of the catalytic function of the RNA hammerhead ribozyme, a three-dimensional model is essential but neither a crystal nor a solution structure has been available. Fluorescence resonance energy transfer (FRET) was used to study the structure of the ribozyme in solution in order to establish the relative spatial orientation of the three constituent Watson-Crick base-paired helical segments. Synthetic constructs were labeled with the fluorescence donor (5-carboxyfluorescein) and acceptor (5-carboxytetramethylrhodamine) located at the ends of the strands constituting the ribozyme molecule. The acceptor helix in helix pairs I and III and in II and III was varied in length from 5 to 11 and 5 to 9 base pairs, respectively, and the FRET efficiencies were determined and correlated with a reference set of labeled RNA duplexes. The FRET efficiencies were predicted on the basis of vector algebra analysis, as a function of the relative helical orientations in the ribozyme constructs, and compared with experimental values. The data were consistent with a Y-shaped arrangement of the ribozyme with helices I and II in close proximity and helix III pointing away. These orientational constraints were used for molecular modeling of a three-dimensional structure of the complete ribozyme.
About this Structure
1RMN is a Protein complex structure of sequences from [1]. Full crystallographic information is available from OCA.
Reference
A three-dimensional model for the hammerhead ribozyme based on fluorescence measurements., Tuschl T, Gohlke C, Jovin TM, Westhof E, Eckstein F, Science. 1994 Nov 4;266(5186):785-9. PMID:7973630
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