title: Regulation of SV40 large T-antigen stability by reversible acetylation.
abstract: Reversible acetylation on protein lysine residues has been shown to regulate the function of both nuclear proteins such as histones and p53 and cytoplasmic proteins such as alpha-tubulin. To identify novel acetylated proteins, we purified several proteins by the affinity to an anti-acetylated-lysine antibody from cells treated with trichostatin A (TSA). Among the proteins identified, here we report acetylation of the SV40 large T antigen (T-Ag). The acetylation site was determined to be lysine-697, which is located adjacent to the C-terminal Cdc4 phospho-degron (CPD). Overexpression of the CBP acetyltransferase acetylated T-Ag, whereas HDAC1, HDAC3 and SIRT1 bound and deacetylated T-Ag. The acetylation and deacetylation occurred independently of p53, a binding partner of T-Ag, but the acetylation was enhanced in the presence of p53. T-Ag in the cells treated with TSA and NA or the acetylation mimic mutant (K697Q) became unstable in COS-7 cells, suggesting that acetylation regulates stability of T-Ag. Indeed, NIH3T3 cells stably expressing K697Q showed decreased anchorage-independent growth compared with those expressing wild type or the K697R mutant. These results demonstrate that acetylation destabilizes T-Ag and regulates the transforming activity of T-Ag in NIH3T3 cells.
authors.affiliation: Chemical Genetics Laboratory, RIKEN, Hirosawa 2-1, Wako, Saitama, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Shimazu
authors.forename: Y
authors.initials: Y
authors.lastname: Komatsu
authors.forename: K I
authors.initials: KI
authors.lastname: Nakayama
authors.forename: H
authors.initials: H
authors.lastname: Fukazawa
authors.forename: S
authors.initials: S
authors.lastname: Horinouchi
authors.forename: M
authors.initials: M
authors.lastname: Yoshida
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D006655:Histone Deacetylases / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D056564:Sirtuin 1; D037761:Sirtuins / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000952:Antigens, Polyomavirus Transforming; D017931:DNA Primers; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D037761:Sirtuins; D006655:Histone Deacetylases