Multiple Roles for Acetylation in the Interaction of p300 HAT with ATF-2 †

Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, 725 North Wolfe Street, Baltimore, Maryland 21205, USA.
Biochemistry (Impact Factor: 3.02). 08/2007; 46(28):8207-16. DOI: 10.1021/bi7000054
Source: PubMed


The transcriptional coactivator paralogues p300 and CBP contain acetyltransferase domains (HAT) and catalyze the lysine acetylation of histones and other proteins as an important aspect of their functions. Prior studies revealed that the basic leucine zipper domain (b-ZIP) of transcription factor ATF-2 (also called CRE-BP1) can interact with the CBP HAT domain. In this study, we have examined the ATF-2 b-ZIP interaction with the p300 HAT domain and shown that p300 HAT autoacetylation can enhance the binding affinity. Pull-down assays revealed that hyperacetylated p300 HAT is more efficiently retained by immobilized ATF-2 b-ZIP than hypoacetylated p300 HAT. Loop deleted p300 HAT lacking autoacetylation was retained about as well as hyperacetylated p300 HAT, suggesting that the loop and ATF-2 compete for p300 HAT binding. While ATF-2 b-ZIP is a weak inhibitor of hypoacetylated p300 HAT acetylation of a histone H4 peptide, hyperacetylated p300 HAT is much more potently inhibited by ATF-2 b-ZIP. Moreover, we showed that ATF-2 b-ZIP could serve as an acetyltransferase substrate for p300 HAT. Using mass spectrometry, two p300 HAT lysine acetylation sites were mapped in ATF-2 b-ZIP. Immunoprecipitation-Western blot analysis with anti-acetyl-lysine antibody revealed that ATF-2 can undergo reversible acetylation in vivo. Mutational analysis of the two ATF-2 b-ZIP acetylation sites revealed their potential contributions to ATF-2-mediated transcriptional activation. Taken together, these studies suggest multiple roles for protein acetylation in the regulation of transcription by p300/CBP and ATF-2.

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    • "In addition to being phosphorylated, ATF2 is also acetylated on Lys357 and Lys374 by p300/CREB-binding protein (CBP, also known as CREBBP), which contributes to its transcriptional activity (Karanam et al., 2007). Binding of ATF2 suppresses the acetyltransferase activity of the transcriptional coactivator p300/CBP. "
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    • "Both p300 and ATF-2 have HAT activity [34, 35]. It was recently shown that p300 acetylates ATF-2 protein in vitro at lysine residues 357 and 374 and that ATF-2 is essential for the acetylation of histones H4 and H2B in vivo [36, 37]. We found that acetylation by p300 is inhibited in a dose-dependent manner by JDP2, when added exogenously. "
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    • "The p300 HAT domain, like that of CBP, acetylates itself and several other domains in p300 in vitro (21–24). Although autoacetylation, so far, has not been studied to a similar extent as p300 acetylation of other proteins, recent studies report that autoacetylation regulates p300 acetyltransferase activity on histone tails and other substrates, which modulates protein–protein interactions and transcription (22,25). Interestingly, the APC/C (anaphase-promoting complex/cyclosome) subunits APC5 and APC7 (26), and GAPDH (glyceraldehydes-3-phosphate dehydrogenase) (27) have been reported to stimulate p300 autoacetylation by directly interacting with p300. "
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