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Assays of nucleosome assembly and the inhibition of histone acetyltransferase activity

External protocol Created on 03 May 2014

Authors

Takahito Yamasaki, Takehide Murata, Chunyuan Jin, Kohsuke Kato, Michiya Noguchi, Koji Nakade, Jianzhi Pan, Kyousuke Nagata, and Kazunari Yokoyama

Summary

Chromatin is a dynamic structure that can adopt markedly different conformations. During transcription, different conformations of chromatin act as important regulatory switches. A novel type of cellular complex, designated INHAT (inhibitor of histone acetyltransferases), was isolated recently and was shown to inhibit the HAT activity of p300/CBP and PCAF by binding to histones, preventing them from serving as substrates for acetyltransferases. This complex was demonstrated, initially, to have nucleosome-assembly activity. In order to examine whether other factors might have both HAT-inhibitory and nucleosome-assembly activities, similar to those of INHATs, we developed methods for measuring the inhibition of HAT activity and nucleosome-assembly activity in vitro and in vivo. Our methodology is particularly useful for measuring the histone-chaperone activity of specific proteins.

Further details

The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.

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