Tracking enzymatic steps of DNA topoisomerases using single-molecule micromanipulation

Strick, T. R., Charvin, G., Dekker, N. H., Allemand, J. F., Bensimon, D., Croquette, V. (June 2002) Tracking enzymatic steps of DNA topoisomerases using single-molecule micromanipulation. Comptes Rendus Physique, 3 (5). pp. 595-618. ISSN 1631-0705

URL: http://www.sciencedirect.com/science/article/pii/S...
DOI: 10.1016/S1631-0705(02)01347-6

Abstract

In this article, we describe single-molecule assays using magnetic traps and we applied these assays to topoisomerase enzymes which unwind and disentangle DNA molecules. First, the elasticity of single DNA molecule is characterized using the magnetic trap. We show that a twisting constraint may be easily applied and that its effect upon DNA may be measured accurately. Then we describe how the topoisomerase activity may be observed at the single-molecule level giving direct access to the important biological parameters of the enzyme such as velocity and processivity. Furthermore, individual cycles of unwinding can be observed in real time. This permits an accurate characterization of the enzyme's biochemical cycle. The data treatment required to identify and analyze individual topoisomerization cycles will be presented in detail. This analysis is applicable to a wide variety of molecular motors.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > topoisomerase
CSHL Authors:
Communities: CSHL labs > Strick lab
Depositing User: Matt Covey
Date: June 2002
Date Deposited: 30 Oct 2013 20:26
Last Modified: 30 Oct 2013 20:26
URI: http://repository.cshl.edu/id/eprint/28796

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