Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.

Tip60 (KAT5) is a histone acetyltransferase (HAT enzyme) involved in multiple cellular processes including transcriptional regulation, DNA damage repair and cell signalling. In prostate cancer, aggressive cases over-express Tip60 which functions as an androgen receptor co-activator via direct acetyl...

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Main Authors: Kelly Coffey, Timothy J Blackburn, Susan Cook, Bernard T Golding, Roger J Griffin, Ian R Hardcastle, Lorraine Hewitt, Kety Huberman, Hesta V McNeill, David R Newell, Celine Roche, Claudia A Ryan-Munden, Anna Watson, Craig N Robson
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0045539&type=printable
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author Kelly Coffey
Timothy J Blackburn
Susan Cook
Bernard T Golding
Roger J Griffin
Ian R Hardcastle
Lorraine Hewitt
Kety Huberman
Hesta V McNeill
David R Newell
Celine Roche
Claudia A Ryan-Munden
Anna Watson
Craig N Robson
author_facet Kelly Coffey
Timothy J Blackburn
Susan Cook
Bernard T Golding
Roger J Griffin
Ian R Hardcastle
Lorraine Hewitt
Kety Huberman
Hesta V McNeill
David R Newell
Celine Roche
Claudia A Ryan-Munden
Anna Watson
Craig N Robson
author_sort Kelly Coffey
collection DOAJ
description Tip60 (KAT5) is a histone acetyltransferase (HAT enzyme) involved in multiple cellular processes including transcriptional regulation, DNA damage repair and cell signalling. In prostate cancer, aggressive cases over-express Tip60 which functions as an androgen receptor co-activator via direct acetylation of lysine residues within the KLKK motif of the receptor hinge region. The purpose of this study was to identify and characterise a Tip60 acetylase inhibitor. High-throughput screening revealed an isothiazole that inhibited both Tip60 and p300 HAT activity. This substance (initially identified as 4-methyl-5-bromoisothiazole) and other isothiazoles were synthesised and assayed against Tip60. Although an authentic sample of 4-methyl-5-bromoisothiazole was inactive against Tip60, in an in vitro HAT assay, 1,2-bis(isothiazol-5-yl)disulfane (NU9056) was identified as a relatively potent inhibitor (IC(50) 2 µM). Cellular activity was confirmed by analysis of acetylation of histone and non-histone proteins in a prostate cancer cell line model. NU9056 treatment inhibited cellular proliferation in a panel of prostate cancer cell lines (50% growth inhibition, 8-27 µM) and induced apoptosis via activation of caspase 3 and caspase 9 in a concentration- and time-dependent manner. Also, decreased androgen receptor, prostate specific antigen, p53 and p21 protein levels were demonstrated in response to treatment with NU9056. Furthermore, pre-treatment with NU9056 inhibited both ATM phosphorylation and Tip60 stabilization in response to ionising radiation. Based on the activity of NU9056 and the specificity of the compound towards Tip60 relative to other HAT enzymes, these chemical biology studies have identified Tip60 as a potential therapeutic target for the treatment of prostate cancer.
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spelling doaj-art-ad0f8a29724847c78bbacae4be0802772025-08-20T02:05:31ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01710e4553910.1371/journal.pone.0045539Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.Kelly CoffeyTimothy J BlackburnSusan CookBernard T GoldingRoger J GriffinIan R HardcastleLorraine HewittKety HubermanHesta V McNeillDavid R NewellCeline RocheClaudia A Ryan-MundenAnna WatsonCraig N RobsonTip60 (KAT5) is a histone acetyltransferase (HAT enzyme) involved in multiple cellular processes including transcriptional regulation, DNA damage repair and cell signalling. In prostate cancer, aggressive cases over-express Tip60 which functions as an androgen receptor co-activator via direct acetylation of lysine residues within the KLKK motif of the receptor hinge region. The purpose of this study was to identify and characterise a Tip60 acetylase inhibitor. High-throughput screening revealed an isothiazole that inhibited both Tip60 and p300 HAT activity. This substance (initially identified as 4-methyl-5-bromoisothiazole) and other isothiazoles were synthesised and assayed against Tip60. Although an authentic sample of 4-methyl-5-bromoisothiazole was inactive against Tip60, in an in vitro HAT assay, 1,2-bis(isothiazol-5-yl)disulfane (NU9056) was identified as a relatively potent inhibitor (IC(50) 2 µM). Cellular activity was confirmed by analysis of acetylation of histone and non-histone proteins in a prostate cancer cell line model. NU9056 treatment inhibited cellular proliferation in a panel of prostate cancer cell lines (50% growth inhibition, 8-27 µM) and induced apoptosis via activation of caspase 3 and caspase 9 in a concentration- and time-dependent manner. Also, decreased androgen receptor, prostate specific antigen, p53 and p21 protein levels were demonstrated in response to treatment with NU9056. Furthermore, pre-treatment with NU9056 inhibited both ATM phosphorylation and Tip60 stabilization in response to ionising radiation. Based on the activity of NU9056 and the specificity of the compound towards Tip60 relative to other HAT enzymes, these chemical biology studies have identified Tip60 as a potential therapeutic target for the treatment of prostate cancer.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0045539&type=printable
spellingShingle Kelly Coffey
Timothy J Blackburn
Susan Cook
Bernard T Golding
Roger J Griffin
Ian R Hardcastle
Lorraine Hewitt
Kety Huberman
Hesta V McNeill
David R Newell
Celine Roche
Claudia A Ryan-Munden
Anna Watson
Craig N Robson
Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.
PLoS ONE
title Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.
title_full Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.
title_fullStr Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.
title_full_unstemmed Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.
title_short Characterisation of a Tip60 specific inhibitor, NU9056, in prostate cancer.
title_sort characterisation of a tip60 specific inhibitor nu9056 in prostate cancer
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0045539&type=printable
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