E2F Family Members Are Differentially Regulated by Reversible Acetylation

Giuseppe Marzio,Christian Wagener,Maria Ines Gutierrez,Peter Cartwright,Kristian Helin,Mauro Giacca

doi:10.1074/jbc.275.15.10887

Giuseppe Marzio, Christian Wagener + Show 4 more

Open Access

https://doi.org/10.1074/jbc.275.15.10887

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Abstract

The six members of the E2F family of transcription factors play a key role in the control of cell cycle progression by regulating the expression of genes involved in DNA replication and cell proliferation. E2F-1, -2, and -3 belong to a structural and functional subfamily distinct from those of the other E2F family members. Here we report that E2F-1, -2, and -3, but not E2F-4, -5, and -6, associate with and are acetylated by p300 and cAMP-response element-binding protein acetyltransferases. Acetylation occurs at three conserved lysine residues located at the N-terminal boundary of their DNA binding domains. Acetylation of E2F-1 in vitro and in vivo markedly increases its binding affinity for a consensus E2F DNA-binding site, which is paralleled by enhanced transactivation of an E2F-responsive promoter. Acetylation of E2F-1 can be reversed by histone deacetylase-1, indicating that reversible acetylation is a mechanism for regulation also of non-histone proteins.

Highlights

Ordered progression through the G1 and S phases of the eukaryotic cell cycle is tightly coupled to the expression of genes whose products are involved in cell growth and DNA replication
A Nuclear Histone Acetyltransferase (HAT) Acetylates E2F Family Members—A current model to explain the role of p300/ cAMP-response element-binding protein (CBP) histone acetyltransferase (HAT) and pRB-associated histone deacetylase-1 (HDAC-1) in the regulation of E2F transactivation predicts that recruitment of these proteins alters the nucleosomes assembled at the promoters of E2Fresponsive genes
The E2F family of transcription factors plays a critical role in the regulation of the cell proliferation by controlling transcription of several genes, whose timely expression is necessary for the ordered progression through the cell cycle

Summary

The abbreviations used are

Retinoblastoma protein; CBP, cAMP-response element-binding protein; HAT, histone acetyltransferase; HDAC-1, histone deacetylase-1; GST, glutathione S-transferase; PAGE, polyacrylamide gel electrophoresis; aa, amino acid; BSA, bovine serum albumin. Few non-histone proteins have recently been discovered to be targets for acetylation by HATs, including TFIIE␤ and TFIIF [34], p53 [35], GATA-1 [36], and the erythroid Kruppel-like factor [37] For these proteins, acetylation is specific for lysine residues. We show that acetylation of E2F-1 can be reversed by HDAC-1, providing for the first time evidence that non-histone proteins are substrates for histone deacetylase enzymes

EXPERIMENTAL PROCEDURES

RESULTS

DISCUSSION