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Live-cell studies of p300/CBP histone acetyltransferase activity and inhibition.

ABSTRACT: Histone acetyltransferase enzymes (HATs) are important therapeutic targets, but there are few cell-based assays available for evaluating the pharmacodynamics of HAT inhibitors. Here we present the application of a FRET-based reporter, Histac, in live-cell studies of p300/CBP HAT inhibition, by both genetic and pharmacologic disruption. shRNA knockdown of p300/CBP led to increased Histac FRET, thus suggesting a role for p300/CBP in the acetylation of the histone H4 tail. Additionally, we describe a new p300/CBP HAT inhibitor, C107, and show that it can also increase cellular Histac FRET. Taken together, these studies provide a live-cell strategy for identifying and evaluating p300/CBP inhibitors.

SUBMITTER: Dancy BM

PROVIDER: S-EPMC3517098 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature

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Live-cell studies of p300/CBP histone acetyltransferase activity and inhibition.

Dancy Beverley M BM Crump Nicholas T NT Peterson Daniel J DJ Mukherjee Chandrani C Bowers Erin M EM Ahn Young-Hoon YH Yoshida Minoru M Zhang Jin J Mahadevan Louis C LC Meyers David J DJ Boeke Jef D JD Cole Philip A PA

Chembiochem : a European journal of chemical biology 20120907 14

Histone acetyltransferase enzymes (HATs) are important therapeutic targets, but there are few cell-based assays available for evaluating the pharmacodynamics of HAT inhibitors. Here we present the application of a FRET-based reporter, Histac, in live-cell studies of p300/CBP HAT inhibition, by both genetic and pharmacologic disruption. shRNA knockdown of p300/CBP led to increased Histac FRET, thus suggesting a role for p300/CBP in the acetylation of the histone H4 tail. Additionally, we describe ...[more]

PMID: 22961914

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