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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    • "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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    ABSTRACT: Transcription factor, Jun dimerization protein 2 (JDP2), binds directly to histones and DNAs and then inhibits the p300-mediated acetylation both of core histones and of reconstituted nucleosomes that contain JDP2 recognition DNA sequences. JDP2 plays a key role as a repressor of adipocyte differentiation by regulation of the expression of the gene C/EBPδ via inhibition of histone acetylation. Moreover, JDP2-deficient mouse embryonic fibroblasts (JDP2−/− MEFs) are resistant to replicative senescence. JDP2 inhibits the recruitment of polycomb repressive complexes (PRC1 and PRC2) to the promoter of the gene encoding p16Ink4a, resulting from the inhibition of methylation of lysine 27 of histone H3 (H3K27). Therefore, it seems that chromatin-remodeling factors, including the PRC complex controlled by JDP2, may be important players in the senescence program. The novel mechanisms that underline the action of JDP2 in inducing cellular senescence and suppressing adipocyte differentiation are reviewed.
    BioMed Research International 01/2011; 2011(6):569034. DOI:10.1155/2011/569034 · 2.71 Impact Factor
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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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    ABSTRACT: MAML1 is a transcriptional coregulator originally identified as a Notch coactivator. MAML1 is also reported to interact with other coregulator proteins, such as CDK8 and p300, to modulate the activity of Notch. We, and others, previously showed that MAML1 recruits p300 to Notch-regulated genes through direct interactions with the DNA-CSL-Notch complex and p300. MAML1 interacts with the C/H3 domain of p300, and the p300-MAML1 complex specifically acetylates lysines of histone H3 and H4 tails in chromatin in vitro. In this report, we show that MAML1 potentiates p300 autoacetylation and p300 transcriptional activation. MAML1 directly enhances p300 HAT activity, and this coincides with the translocation of MAML1, p300 and acetylated histones to nuclear bodies.
    Nucleic Acids Research 04/2009; 37(9):2996-3006. DOI:10.1093/nar/gkp163 · 9.11 Impact Factor
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    • "A cell could specify different transcriptional programs by controlling the dynamic ratio of ac-p300/p300 through SIRT2. Consistent with this idea, p300 isolated from cells treated with HDAC inhibitors interacts more tightly with p53 (Stiehl et al., 2007) and ATF-2 preferentially interacts with ac-p300 in vitro (Karanam et al., 2007). The dynamics of p300 autoacetylation and deacetylation by SIRT2 provide another level of precision in the fine-tuning of gene regulation programs. "
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    ABSTRACT: Autoacetylation of the p300 histone acetyltransferase controls the transition between VP16-mediated chromatin acetylation and preinitiation complex (PIC) assembly. Currently, it is unknown if and how autoacetylated p300 is deacetylated. We found that the NAD(+)-dependent histone deacetylase SIRT2 deacetylates p300 in vitro and in cells. SIRT2 deacetylates lysine residues in the catalytic domain of p300 and restores binding of p300 to the PIC. RNAi-mediated depletion or chemical inhibition of SIRT2 in cells results in accumulation of acetylated p300. The altered ac-p300/p300 ratio in SIRT2-depleted cells results in decreased p300 recruitment to an integrated VP16-responsive gene and inhibition of transcription. We conclude that p300 undergoes a dynamic cycle of autoacetylation and deacetylation.
    Molecular cell 12/2008; 32(3):449-55. DOI:10.1016/j.molcel.2008.09.018 · 14.02 Impact Factor
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