Trichostatin A Induces Transforming Growth Factor β Type II Receptor Promoter Activity and Acetylation of Sp1 by Recruitment of PCAF/p300 to a Sp1·NF-Y Complex

Abstract

Transforming growth factor ␤ type II receptor (T␤RII) is a tumor suppressor gene that can be transcriptionally silenced by histone deacetylases (HDACs) in cancer cells. In this report, we demonstrated the mechanism by which trichostatin A (TSA), an inhibitor of HDAC, induces the expression of T␤RII in human pancreatic cancer cell lines by modulating the transcriptional components that bind a specific DNA region of the T␤RII promoter. This region of the T␤RII promoter possesses Sp1 and NF-Y binding sites in close proximity (located at ؊102 and ؊83, respectively). Treatment of cells with TSA activates the T␤RII promoter in a time-dependent manner through the recruitment of p300 and PCAF into a Sp1⅐NF-Y⅐HDAC complex that binds this DNA element. The recruitment of p300 and PCAF into the complex is associated with a concomitant acetylation of Sp1 and an overall decrease in the amount of HDAC associated with the complex. Transient overexpression of p300 or PCAF potentiated TSA-induced T␤RII promoter activity. The effect of PCAF was dependent on its histone acetyltransferase activity, whereas that of p300 was independent. Stable transfection of PCAF caused an increase in T␤RII mRNA expression, the association of PCAF with T␤RII promoter, and the acetylation of Sp1. Taken together, these results showed that TSA treatment of pancreatic cancer cells leads to transcriptional activation of the T␤RII promoter through modulation of the components of a Sp1⅐NF-Y⅐p300⅐PCAF⅐HDAC-1 multiprotein complex. Moreover, the interaction of NF-Y with the Sp1-associated complex may further explain why this specific Sp1 site mediates transcriptional responsiveness to TSA. TGF-␤1 plays a significant role in the growth inhibition of most normal epithelial and some cancer cells (1). TGF-␤ mediates its biological effects through cell surface receptors known as TGF-␤ type I receptor (T␤RI) and TGF-␤ type II receptor (T␤RII). Its intracellular signaling is initiated upon the selective binding of the active cytokine to the T␤RII homodimer. T␤RII is a ubiquitously expressed and constitutively active serine/threonine kinase. Ligand binding to T␤RII induces the assembly of a heterotetrameric complex consisting of T␤RI and T␤RII. Once the receptor complex is formed, T␤RII phosphorylates and thereby activates the T␤RI serine/threonine kinase. Activation of T␤RI propagates downstream signaling via Smad family proteins. T␤RI directly interacts with and phosphorylates Smad2 and Smad3. These Smads bind Smad4 and then result in the translocation of this complex to the nucleus and modulate TGF-␤-responsive gene expression (2-4).The TGF-␤ signaling pathway is inactivated in many tumors. Loss of negative growth regulation by TGF-␤ affords cells a selective growth advantage associated with decreased dependence of exogenous growth factor and increased tumorigenicity. Frequently, inhibition of TGF-␤ signaling occurs by either abolition of the function of a common mediator, Smad4, or interference with T␤RII function (5, 6). Smad4 and T␤RII are tumor suppressor gene...

Dynamic transcriptional regulatory complexes including BORIS, CTCF and Sp1 modulate NY-ESO-1 expression in lung cancer cells

“…In addition, post-translational modifications of Sp1 may also augment interaction of this transcription factor with the NY-ESO-1 promoter. Several recent studies indicate that HDAC inhibitors mediate acetylation of Sp1, and enhance binding of Sp1 to target recognition sequences (Ryu et al, 2003;Huang et al, 2005). Finally, our data do not exclude the possibility that micro-RNAs induced by HDAC inhibition (Scott et al, 2006) augment expression of NY-ESO-1.…”

Histone deacetylase inhibitors induce FPGS mRNA expression and intracellular accumulation of long-chain methotrexate polyglutamates in childhood acute lymphoblastic leukemia: implications for combination therapy

“…Consequently, as observed, treatment with HDACIs would alter the FPGS promoter acetylation status, modify its chromatin structure and lead to increased FPGS gene transcription. Similar interactions between NFY, Sp1 and HDAC1 have been reported in other TATA-less promoters such as the transforming growth factor-b type-II receptor (TbRII) 55 and the HDAC1 promoter. 56 In the TbRII gene, HDAC activity selectively blocked the ability of Sp1-and NFY-binding sites to activate the TbRII promoter transcription and allow the PCAF protein to associate to the NFY complex to bind to the CCAAT-box in the TbRII promoter.…”

The histone deacetylase inhibitor trichostatin A induces GADD45γ expression via Oct and NF-Y binding sites

“…Role of co-repressors and co-activators in the regulation of the murine GADD45g promoter NF-Y and Oct-1 can interact with HAT-containing coactivator complexes and HDAC-containing co-repressor complexes (Nagy et al, 1997;Mantovani, 1999;Kakizawa et al, 2001;Park et al, 2002;Peng and Jahroudi, 2003;Huang et al, 2005). We therefore assessed the contribution of these factors to GADD45g promoter activity regulation.…”

“…NF-Y recruits the histone acetylase PCAF to promoters (Park et al, 2002;Huang et al, 2005). The participation of PCAF in the regulation of the GADD45g promoter was addressed by co-transfecting a PCAF expression plasmid with the À818 to þ 15 promoter reporter plasmid.…”

“…Since Oct-1 and NF-Y can interact with HATcontaining co-activator complexes and HDAC-containing co-repressor complexes (Nagy et al, 1997;Mantovani, 1999;Kakizawa et al, 2001;Park et al, 2002;Peng and Jahroudi, 2003;Huang et al, 2005), we studied the possible role of SMRT, HDAC1, and PCAF in the regulation of the murine GADD45g promoter. SMRT or HDAC1, but not HDAC3, ectopic expression reduced GADD45g promoter activity, while forced expression of PCAF increased it.…”