Regulation of the p300 HAT domain via a novel activation loop

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The transcriptional coactivator p300 is a histone acetyltransferase (HAT) whose function is critical for regulating gene expression in mammalian cells. However, the molecular events that regulate p300 HAT activity are poorly understood. We evaluated autoacetylation of the p300 HAT protein domain to determine its function. Using expressed protein ligation, the p300 HAT protein domain was generated in hypoacetylated form and found to have reduced catalytic activity. This basal catalytic rate was stimulated by autoacetylation of several key lysine sites within an apparent activation loop motif. This post-translational modification and catalytic regulation of p300 HAT activity is conceptually analogous to the activation of most protein kinases by autophosphorylation. We therefore propose that this autoregulatory loop could influence the impact of p300 on a wide variety of signaling and transcriptional events.

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Figure 1: Purification and partial proteolysis of p300(1195–1673) HAT domain.
Figure 2: Expression and purification of semisynthetic p300 HAT domain by expressed protein ligation.
Figure 3: Autoacetylation of p300.
Figure 4: A proteolytically sensitive loop region that regulates p300 HAT activity.
Figure 5: Role of p300 autoacetylation in vivo.

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This work was supported by grants from the US National Institutes of Health to P.A.C. and J.W. and from the Ellison Medical Foundation to P.A.C., by a Canadian Institutes for Health Research postdoctoral fellowship to P.R.T., and by grants from AIRC, MURST-Cofin and MURST-FIRB to M.L. W.A. was supported by a National Research Fellowship Award. We thank C. Wolberger, M. Ott and J. Boeke for helpful discussions and for reagents. We thank N. Rust for technical assistance. We thank D. Leahy, R. Alani and J. Liu for comments on the manuscript.

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Correspondence to Philip A Cole.

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Q.G. works for Cell Signaling Technology Inc. and could benefit by selling antibody. R.J.C. is a paid consultant for Shimadzu Co. and has licensed his invention to the company.

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Thompson, P., Wang, D., Wang, L. et al. Regulation of the p300 HAT domain via a novel activation loop. Nat Struct Mol Biol 11, 308–315 (2004) doi:10.1038/nsmb740

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