Acetylation of p53 at Lysine 373/382 by the Histone Deacetylase Inhibitor Depsipeptide Induces Expression of p21^Waf1/Cip1

Abstract

Generally, histone deacetylase (HDAC) inhibitor-induced p21Waf1/Cip1 expression is thought to be p53 independent. Here we found that an inhibitor of HDAC, depsipeptide (FR901228), but not trichostatin A (TSA), induces p21 Waf1/Cip1 expression through both p53 and Sp1/Sp3 pathways in A549 cells (which retain wild-type p53). This is demonstrated by measuring relative luciferase activities of p21 promoter constructs with p53 or Sp1 binding site mutagenesis and was further confirmed by transfection of wild-type p53 into H1299 cells (p53 null). That p53 was acetylated after depsipeptide treatment was tested by sequential immunoprecipitation/Western immunoblot analysis with anti-acetylated lysines and anti-p53 antibodies. The acetylated p53 has a longer half-life due to a significant decrease in p53 ubiquitination. Further study using site-specific antiacetyllysine antibodies and transfection of mutated p53 vectors (K319/K320/K321R mutated and K373R/K382R mutations) into H1299 cells revealed that depsipeptide specifically induces p53 acetylation at K373/K382, but not at K320. As assayed by coimmunoprecipitation, the K373/K382 acetylation is accompanied by a recruitment of p300, but neither CREB-binding protein (CBP) nor p300/CBP-associated factor (PCAF), to the p53 C terminus. Furthermore, activity associated with the binding of the acetylated p53 at K373/K382 to the p21 promoter as well as p21Waf1/Cip1 expression is significantly increased after depsipeptide treatment, as tested by chromatin immunoprecipitations and Western blotting, respectively. In addition, p53 acetylation at K373/K382 is confirmed to be required for recruitment of p300 to the p21 promoter, and the depsipeptide-induced p53 acetylation at K373/K382 is unlikely to be dependent on p53 phosphorylation at Ser15, Ser20, and Ser392 sites. Our data suggest that p53 acetylation at K373/K382 plays an important role in depsipeptide-induced p21 Waf1/Cip1 expression.

HDAC4 represses p21WAF1/Cip1 expression in human cancer cells through a Sp1-dependent, p53-independent mechanism

“…HDAC4 represses p21 WAF1/Cip1 in cancer cells in vitro in a Sp1-dependent manner Although most reports demonstrated that HDAC inhibitors induced p21 WAF1/Cip1 expression in an Sp1/Sp3-dependent manner Sowa et al, 1997Sowa et al, , 1999Han et al, 2001;Wilson et al, 2006), a role for p53 in HDAC-associated p21 WAF1/Cip1 expression has also been reported in several human cancer cell lines (Lagger et al, 2003;Luo et al, 2004;Roy et al, 2005;Zhao et al, 2006).…”

“…To date, the most common model for p21 WAF1/Cip1 repression in cancer cells is the recruitment of HDACs by Sp1/-Sp3 transcription factors in the proximal promoter region encompassing the Sp1/Sp3 binding sites, which in turn repress p21 WAF1/Cip1 transcription by deacetylating histones H3 and H4. However, a role for p53 in HDAC-associated p21 WAF1/Cip1 expression has also been reported in several human cancer cell lines (Lagger et al, 2003;Luo et al, 2004;Roy et al, 2005;Zhao et al, 2006).…”

Mule determines the apoptotic response to HDAC inhibitors by targeted ubiquitination and destruction of HDAC2

“…p53 acetylation is removed by HDACs. HDAC1 has been shown to deacetylate p53 at Lys 373/382 to induce p21 expression (Zhao et al 2006). p21 can also be activated by HDACis through p53-independent pathways (Gartel and Tyner 1999;Ocker and Schneider-Stock 2007).…”

Cutaneous T‐cell lymphoma: 2011 update on diagnosis, risk‐stratification, and management

“…The 18 human HDACs may be classified as either zinc-or NAD 1 -dependent and further subclassified into class I (HDAC1, 2, 3, and 8), class II (HDAC4, 5, 6, 7, 9, and 10), class III (including NAD 1 -dependent sirtuins), and class IV (HDAC11) HDACs. As HDACs regulate a wide variety of processes involved in carcinogenesis, multiple mechanisms may explain the clinical activity of HDAC inhibitors [249,250], including altered gene expression of cell-cycle and apoptotic regulatory proteins [251][252][253][254][255], acetylation of nonhistone proteins regulating cell growth and survival [256][257][258][259], angiogenesis [260,261], aggresome formation [262], and DNA repair [263]. In addition, HDAC inhibitors may have important effects on the tumor microenvironment via reactive oxygen species [264,265], enhanced antigen presentation [266] and downregulation of immunomodulatory cytokines, like IL-10 [267].…”