Document Detail


The effect of physical form of DNA on exonucleaseIII activity revealed by single-molecule observations.
MedLine Citation:
PMID:  18561002     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Single-molecule studies have revealed molecular behaviors usually hidden in the ensemble and time averaging of bulk experiments. Single-molecule measurement that can control physical form of individual DNA molecules is a powerful method to obtain new knowledge about correlation between DNA-tension and enzyme activity. Here we study the effect of physical form of DNA on exonucleaseIII (ExoIII) reaction. ExoIII has a double-stranded DNA specific 3'-->5' exonuclease activity and the digestion is distributive. We observed the ExoIII digestion of individual stretched DNA molecules from the free ends. The sequentially captured photographs demonstrated that the digested DNA molecule linearly shortened with the reaction time. We also carried out the single-molecule observation under random coiled form by pausing the buffer flow. The digestion rates obtained from both single-molecule experiments showed that the digestion rate under the stretched condition was two times higher than the random coiled condition. The correlation between physical form of DNA and digestion rate of ExoIII was clearly demonstrated by single-molecule observations.
Authors:
Hirofumi Kurita; Ken Torii; Hachiro Yasuda; Kazunori Takashima; Shinji Katsura; Akira Mizuno
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-06-17
Journal Detail:
Title:  Journal of fluorescence     Volume:  19     ISSN:  1053-0509     ISO Abbreviation:  J Fluoresc     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2009-02-05     Completed Date:  2009-05-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9201341     Medline TA:  J Fluoresc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  33-40     Citation Subset:  IM    
Affiliation:
Department of Ecological Engineering, Toyohashi University of Technology, Aichi, 441-8580, Japan.
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MeSH Terms
Descriptor/Qualifier:
DNA / chemistry*,  metabolism*
Enzyme Activation
Enzymes, Immobilized / chemistry,  metabolism
Exodeoxyribonucleases / analysis,  metabolism*
Fluorescence
Time Factors
Chemical
Reg. No./Substance:
0/Enzymes, Immobilized; 9007-49-2/DNA; EC 3.1.-/Exodeoxyribonucleases; EC 3.1.11.2/exodeoxyribonuclease III

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