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Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium

[Image: see text] Pyrvinium was identified as the first small molecule inhibitor of the androgen receptor (AR) DNA-binding domain (DBD). It was also among the first small molecules shown to directly inhibit the activity of AR splice variants (ARVs), which has important clinical implications in the t...

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Autores principales: Pal, Sumanta K., Tew, Ben Yi, Lim, Minyoung, Stankavich, Brittany, He, Miaoling, Pufall, Miles, Hu, Weidong, Chen, Yuan, Jones, Jeremy O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2019
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6410682/
https://www.ncbi.nlm.nih.gov/pubmed/30873507
http://dx.doi.org/10.1021/acsomega.8b03205
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author Pal, Sumanta K.
Tew, Ben Yi
Lim, Minyoung
Stankavich, Brittany
He, Miaoling
Pufall, Miles
Hu, Weidong
Chen, Yuan
Jones, Jeremy O.
author_facet Pal, Sumanta K.
Tew, Ben Yi
Lim, Minyoung
Stankavich, Brittany
He, Miaoling
Pufall, Miles
Hu, Weidong
Chen, Yuan
Jones, Jeremy O.
author_sort Pal, Sumanta K.
collection PubMed
description [Image: see text] Pyrvinium was identified as the first small molecule inhibitor of the androgen receptor (AR) DNA-binding domain (DBD). It was also among the first small molecules shown to directly inhibit the activity of AR splice variants (ARVs), which has important clinical implications in the treatment of castration-resistant prostate cancer. Important questions about pyrvinium’s mechanism of action remain. Here, we demonstrate through mutational analysis that amino acids 609 and 612 are important for pyrvinium action. Nuclear magnetic resonance demonstrates a specific interaction between a soluble pyrvinium derivative and the AR DBD homodimer–DNA complex. Chromatin immunoprecipitation and electrophoretic mobility shift assay experiments demonstrate that, despite an interaction with this complex, pyrvinium does not alter the DNA-binding kinetics in either assay. AR immunoprecipitation followed by mass spectrometry was used to identify proteins whose interaction with AR is altered by pyrvinium. Several splicing factors, including DDX17, had reduced interactions with AR in the presence of pyrvinium. RNA sequencing of prostate cancer cells treated with pyrvinium demonstrated changes in splicing, as well as in several other pathways. However, pyrvinium did not alter the levels of ARVs in several prostate cancer cell lines. Taken together, our new data pinpoint the direct interaction between pyrvinium and AR DBD and shed light on the mechanism by which it inhibits AR transcriptional activity.
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spelling pubmed-64106822019-03-12 Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium Pal, Sumanta K. Tew, Ben Yi Lim, Minyoung Stankavich, Brittany He, Miaoling Pufall, Miles Hu, Weidong Chen, Yuan Jones, Jeremy O. ACS Omega [Image: see text] Pyrvinium was identified as the first small molecule inhibitor of the androgen receptor (AR) DNA-binding domain (DBD). It was also among the first small molecules shown to directly inhibit the activity of AR splice variants (ARVs), which has important clinical implications in the treatment of castration-resistant prostate cancer. Important questions about pyrvinium’s mechanism of action remain. Here, we demonstrate through mutational analysis that amino acids 609 and 612 are important for pyrvinium action. Nuclear magnetic resonance demonstrates a specific interaction between a soluble pyrvinium derivative and the AR DBD homodimer–DNA complex. Chromatin immunoprecipitation and electrophoretic mobility shift assay experiments demonstrate that, despite an interaction with this complex, pyrvinium does not alter the DNA-binding kinetics in either assay. AR immunoprecipitation followed by mass spectrometry was used to identify proteins whose interaction with AR is altered by pyrvinium. Several splicing factors, including DDX17, had reduced interactions with AR in the presence of pyrvinium. RNA sequencing of prostate cancer cells treated with pyrvinium demonstrated changes in splicing, as well as in several other pathways. However, pyrvinium did not alter the levels of ARVs in several prostate cancer cell lines. Taken together, our new data pinpoint the direct interaction between pyrvinium and AR DBD and shed light on the mechanism by which it inhibits AR transcriptional activity. American Chemical Society 2019-02-01 /pmc/articles/PMC6410682/ /pubmed/30873507 http://dx.doi.org/10.1021/acsomega.8b03205 Text en Copyright © 2019 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Pal, Sumanta K.
Tew, Ben Yi
Lim, Minyoung
Stankavich, Brittany
He, Miaoling
Pufall, Miles
Hu, Weidong
Chen, Yuan
Jones, Jeremy O.
Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium
title Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium
title_full Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium
title_fullStr Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium
title_full_unstemmed Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium
title_short Mechanistic Investigation of the Androgen Receptor DNA-Binding Domain Inhibitor Pyrvinium
title_sort mechanistic investigation of the androgen receptor dna-binding domain inhibitor pyrvinium
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6410682/
https://www.ncbi.nlm.nih.gov/pubmed/30873507
http://dx.doi.org/10.1021/acsomega.8b03205
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