Live-Cell Studies of p300/CBP Histone Acetyltransferase Activity and Inhibition

Authors

  • Beverley M. Dancy,

    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    2. Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Nicholas T. Crump,

    1. Nuclear Signalling Laboratory, Department of Biochemistry, Oxford University, South Parks Road, Oxford OX1 3QU (UK)
    Search for more papers by this author
  • Daniel J. Peterson,

    1. Laboratory for Neurocognitive and Imaging Research, Kennedy Krieger Institute, 716 North Broadway, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Chandrani Mukherjee,

    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Erin M. Bowers,

    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Young-Hoon Ahn,

    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Minoru Yoshida,

    1. Chemical Genetics Laboratory, RIKEN Advanced Science Institute, Wako, Saitama 351-0198 (Japan)
    Search for more papers by this author
  • Dr. Jin Zhang,

    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Louis C. Mahadevan,

    1. Nuclear Signalling Laboratory, Department of Biochemistry, Oxford University, South Parks Road, Oxford OX1 3QU (UK)
    Search for more papers by this author
  • Dr. David J. Meyers,

    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Jef D. Boeke,

    1. Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD 21205 (USA)
    Search for more papers by this author
  • Dr. Philip A. Cole

    Corresponding author
    1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    • Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205 (USA)
    Search for more papers by this author

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.

Ancillary