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Updated: Jul 9, 2026
Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo
Published on: January 14, 2016
Arabidopsis HY5 protein functions as a DNA-binding tag for purification and functional immobilization of proteins on
Tatsuya Sawasaki1, Nami Kamura, Satoko Matsunaga
1Cell-Free Science and Technology Research Center, and The Venture Business Laboratory, Ehime University, Matsuyama, Ehime, Japan. sawasaki@eng.ehime-u.ac.jp
Abstract:
Protein microarray is considered to be one of the key analytical tools for high-throughput protein function analysis. Here, we report that the Arabidopsis HY5 functions as a novel DNA-binding tag (DBtag) for proteins. We also demonstrate that the DBtagged proteins could be immobilized and purified on a newly designed agarose/DNA microplate. Furthermore, we show three applications using the microarray: (1) detection of autophosphorylation activity of DBtagged human protein kinases and inhibition of their activity by staurosporine, (2) specific cleavage of DBtagged proteins by a virus protease and caspase 3, and (3) detection of a protein-protein interaction between the DBtagged UBE2N and UBE2v1. Thus, this method may facilitate rapid functional analysis of a wide range of proteins.
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