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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
27
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pubmed:dateCreated |
2004-7-8
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pubmed:abstractText |
The hairpin ribozyme is a minimalist paradigm for studying RNA folding and function. In this enzyme, two domains dock by induced fit to form a catalytic core that mediates a specific backbone cleavage reaction. Here, we have fully dissected its reversible reaction pathway, which comprises two structural transitions (docking/undocking) and a chemistry step (cleavage/ligation), by applying a combination of single-molecule fluorescence resonance energy transfer (FRET) assays, ensemble cleavage assays, and kinetic simulations. This has allowed us to quantify the effects that modifications of essential functional groups remote from the site of catalysis have on the individual rate constants. We find that all ribozyme variants show similar fractionations into effectively noninterchanging molecule subpopulations of distinct undocking rate constants. This leads to heterogeneous cleavage activity as commonly observed for RNA enzymes. A modification at the domain junction additionally leads to heterogeneous docking. Surprisingly, most modifications not only affect docking/undocking but also significantly impact the internal chemistry rate constants over a substantial distance from the site of catalysis. We propose that a network of coupled molecular motions connects distant parts of the RNA with its reaction site, which suggests a previously undescribed analogy between RNA and protein enzymes. Our findings also have broad implications for applications such as the action of drugs and ligands distal to the active site or the engineering of allostery into RNA.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-10438622,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-10554775,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-10715200,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11298439,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11326079,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11406379,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11558994,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11707414,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11983335,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-11987183,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12029135,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12376595,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12581665,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12649489,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12869691,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12883002,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-12947189,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-14527328,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-14691941,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-9153212,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-9545249,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-9667918,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15218105-9836635
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
6
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pubmed:volume |
101
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
10066-71
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
2004
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pubmed:articleTitle |
Single-molecule enzymology of RNA: essential functional groups impact catalysis from a distance.
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pubmed:affiliation |
Department of Chemistry, University of Michigan, Ann Arbor, 48109, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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