A specific lysine in c-Jun is required for transcriptional repression by E1A and is acetylated by p300

Abstract

The adenovirus E1A protein regulates transcription of cellular genes via its interaction with the transcriptional coactivators p300/CBP. The collagenase promoter activated by the c-Jun protein is repressed by E1A. Here we show that E1A repression is specific for c-Jun, as E1A does not repress the collagenase promoter activated by the homologous transcription factor EB1. Using chimeras of c-Jun and EB1, we demonstrate that a 12 amino acid region in the basic region of the c-Jun DNA-binding domain is essential for repression by E1A. Since repression requires the binding of p300 to E1A, we studied the involvement of p300 acetyltransferase activity in the repression mechanism. We demonstrate that c-Jun is acetylated in vivo, and mutational analysis identified Lys271 in the c-Jun basic region to be essential for repression of the collagenase promoter by E1A. In addition, Lys271 is acetylated both in vitro and in vivo. These results suggest that the specific repression of the collagenase promoter by E1A involves acetylation of c-Jun.

FOXO4 Is Acetylated upon Peroxide Stress and Deacetylated by the Longevity Protein hSir2

“…The experiments presented are representative of at least three independent experiments. DB) was in vitro acetylated by a GST-tagged p300 acetyltransferase domain in the presence of [ 14 C]acetyl-CoA (21). Subsequently, the reactions were incubated with GST-tagged hSir2 SIRT1 in the absence or presence of the essential cofactor NAD ϩ (24).…”

“…pcDNA3.1-myc-hSir2 SIRT1 and pRP265-GSThSir2 SIRT1 were created by ligating a BamHI/XbaI fragment from pcDNA3.1-hSir2 SIRT1 into BamHI/XbaI-digested pcDNA3.1-myc and pRP265, respectively. pRc/RSV-CBP-HA and pRP265-GST-p300-HAT were kindly provided by R. Giles (20) and R. Vries (21), respectively. Gal4-DBD-CBP-HA constructs were a kind gift of E. Kalkhoven (22).…”

“…In Vitro and in Vivo Deacetylation-For in vitro deacetylation, 2.5 g GST-FOXO4-DB were incubated with 0.05 Ci [ 14 C]-acetyl-CoA (ICN) and 2.0 g GST-p300-HAT in reaction buffer (50 mM Tris-HCl, pH 8.0, 5 mM EDTA, pH 8.0, 0.5% Nonidet P-40, 150 mM NaCl (21) for 45 min at 30°C. Subsequently, deacetylation buffer (50 mM Tris-HCl, pH 8.8, 4 mM MgCl 2 , 0.2 mM dithiothreitol, 2 M trichostatin A plus 1 mM NAD ϩ if indicated (24) and 2.0 g of GST-tagged wt-hSir2 SIRT1 were added to a final volume of 100 l. Samples were incubated for another 90 min at 37°C, and the reaction was terminated using 5ϫ Laemmli sample buffer.…”

SIRT1 Suppresses Activator Protein-1 Transcriptional Activity and Cyclooxygenase-2 Expression in Macrophages

“…In line with these observations and extended to c-Fos, we found that SIRT1 directly interacted with the bZIP domain of both c-Fos and c-Jun, resulting in a reduction in the p300-induced acetylation of c-Fos and c-Jun. c-Jun has been reported to be acetylated by CBP/p300 at 271 lysine residues within the basic region of bZIP domain (5), which promoted us to investigate the relationships between the acetylation status of the basic region and SIRT1-mediated transcriptional inhibition. Surprisingly, none of the lysine mutants within the basic region of c-Jun affected AP-1 transcriptional activity in our experiments (data not shown).…”

“…In addition to direct binding to cis-acting elements, AP-1 could also interact with other transcriptional regulators, participating in transcriptional regulation, and the bZIP domain contributes to the association with co-activator (3) or co-repressor (4). The transcriptional activity of AP-1 is regulated by the post-translational modification, including acetylation (5). The acetylation modification of transcriptional factors changes the proteinprotein or protein-DNA interaction as an important regulatory mechanism of transcription (6).…”