Of ARD1 in LNCaP cells AR easier to communicate with the PSA or TMPRSS2 promoter, as compared to that in cells without overexpression of ARD1. Together, these results indicated that ARD1 acetyl inducedARD1 for AR target gene transcription. The mechanism of gene transcription JNK Signaling Pathway mediated ARD1 f AR Promotes understanding, we examined the AR level or stability T by ARD1 downregulation. Curiously, reduce ARD1 silencing fact, the level of PSA, but seemed to have no influence on the AR level have. Because ARD1 is an acetyltransferase, we assumed that ARD1 could exert its regulatory activity of AR acetylation. Analysis of mutual co-Immunpr Zipitation of endogenous or exogenous proteins, ARD1 and AR in 293T and LNCaP cells showed that ARD1 physically interacts with AR in vivo and in vitro. With rpern Antique Who found specific acetylation, we find that ARD1 silencing in LNCaP cells inhibits AR acetylation significantly. To determine whether AR ARD1 ARD1-mediated acetylation is specific, we generated a catalytically inactive mutant ARD1 as described above. The expression of mutant ARD1 significantly reduced the level of dead acetylated AR. To the r best Term Was the ARD1 in the acetylation of AR conducted an in vitro assay using purified recombinant acetylation Volll Nts-AR as the substrate, the immunpr Zipitierte WT ARD1 ARD1 or dead, there the enzyme, and acetyl-CoA as coenzyme.We showed that ARD1 acetyl RA directly in vitro, but not dead ARD1. Close Of course, we investigated the m Possible connection between acetylation of ARD1 Ar and Ar target gene transcription. We have shown that the mutant ARD1 dead was not able either to Promotoraktivit t to induce PSA or PSA transactivation in LNCaP cells by testing luciferase reporter or by qRT-PCR, respectively. In addition, reduced dead mutant ARD1 bind the F Demonstrated ability of AR to promoters of PSA by chip analysis.
These results show that the acetylation of AR ARD1 ARD1 necessary for induced AR target gene transcription and tumorigenesis of prostate. Discussion In this study, we show an R The single ARD1as anARregulator in prostate tumorigenesis. We show that the interaction and ARD1 ARD1 AR AR dependent Independent acetylation for transcriptional activation of AR target genes in vivo and in vitro are needed. ARD1 silence in LNCaP cells leads to a strong inhibition of AR target gene transcription, cell proliferation of prostate cancer and xenograft tumor growth. The importance of the AR acetylation of ARD1 is further strengthened by the demonstration that bring silence to express ARD1 ARD1 or mutated AR acetylation reduces Todesf Ll founded and led to reduced interaction of AR-target promoter and expression of genes ARtarget. It has been reported that AR can be acetylated by p300 on lysine residues 630 AR, 632, 633 and AR-ligand-dependent Independent activation. Curiously, p300 silencing that reducing the level of the entire AR, but not the level of the ARD1 acetylated AR. In addition, Arin mutant with three lysine residues were replaced by glutamine is perhaps acetylated by ARD1. These data suggest that AR acetylation site for ARD1 k Nnte different from that of p300. Further analysis is needed to identify the specific acetylation site in AR ARD1 to better fully understand the mechanisms, through the mediation of ARD1 acetyl f AR AR trans To be rdern.