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A DNA immunoprecipitation assay used in quantitative detection of in vitro DNA-protein complex binding

Title
A DNA immunoprecipitation assay used in quantitative detection of in vitro DNA-protein complex binding
Author
채지형
Keywords
DNA-protein interaction; DNA immunoprecipitation; Transcription factor CP2c; DNA-protein binding dynamics; TRANSCRIPTION FACTOR CP2; LATE SV40 FACTOR; ALPHA-GLOBIN GENE; FACTOR LSF; HEPATOCELLULAR-CARCINOMA; GEL-ELECTROPHORESIS; IDENTIFICATION; LOCALIZATION; EXPRESSION; ONCOGENE
Issue Date
2013-10
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE, 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA
Citation
Analytical Biochemistry, 2013, 441(2), P.147-151
Abstract
To begin gene transcription, several transcription factors must bind to specific DNA sequences to form a complex via DNA-protein interactions. We established an in vitro method for specific and sensitive analyses of DNA-protein interactions based on a DNA immunoprecipitation (DIP) method. We verified the accuracy and efficiency of the DIP assay in quantitatively measuring DNA-protein binding using transcription factor CP2c as a model. With our DIP assay, we could detect specific interactions within a DNA-CP2c complex, with reproducible and quantitative binding values. In addition, we were able to effectively measure the changes in DNA-CP2c binding by the addition of a small molecule, FQI1 (factor quinolinone inhibitor 1), previously identified as a specific inhibitor of this binding. To identify a new regulator of DNA-CP2c binding, we analyzed several CP2c binding peptides and found that only one class of peptide severely inhibits DNA-CP2c binding. These data show that our DIP assay is very useful in quantitatively detecting the binding dynamics of DNA-protein complex. Because DNA-protein interaction is very dynamic in different cellular environments, our assay can be applied to the detection of active transcription factors, including promoter occupancy in normal and disease conditions. Moreover, it may be used to develop a targeted regulator of specific DNA-protein interaction. Crown Copyright (c) 2013 Published by Elsevier Inc. All rights reserved.
URI
https://ac.els-cdn.com/S0003269713003151/1-s2.0-S0003269713003151-main.pdf?_tid=01fb5748-3d0a-4b29-bcbe-1bcb83cb6200&acdnat=1520340692_cf6aedfc5079f0ce11fe7eb1db942aebhttp://hdl.handle.net/20.500.11754/44448
ISSN
0003-2697; 1096-0309
DOI
10.1016/j.ab.2013.07.001
Appears in Collections:
CENTER FOR CREATIVE CONVERGENCE EDUCATION[S](창의융합교육원) > ETC
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