In vitro selection of mutant HDM2 resistant to nutlin inhibition

HDM2 binds to the p53 tumour suppressor and targets it for proteosomal degradation. Presently in clinical trials, the small molecule Nutlin-3A competitively binds to HDM2 and abrogates its repressive function. Using a novel in vitro selection methodology, we simulated the emergence of resistance by...

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Main Authors: Wei, Siau Jia., Joseph, Thomas., Sim, Adelene Y. L., Yurlova, Larisa., Zolghadr, Kourosh., Lane, David., Verma, Chandra., Ghadessy, Farid.
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2014
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Online Access:https://hdl.handle.net/10356/100150
http://hdl.handle.net/10220/18644
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0062564
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-1001502023-02-28T17:02:51Z In vitro selection of mutant HDM2 resistant to nutlin inhibition Wei, Siau Jia. Joseph, Thomas. Sim, Adelene Y. L. Yurlova, Larisa. Zolghadr, Kourosh. Lane, David. Verma, Chandra. Ghadessy, Farid. School of Biological Sciences DRNTU::Science::Biological sciences HDM2 binds to the p53 tumour suppressor and targets it for proteosomal degradation. Presently in clinical trials, the small molecule Nutlin-3A competitively binds to HDM2 and abrogates its repressive function. Using a novel in vitro selection methodology, we simulated the emergence of resistance by evolving HDM2 mutants capable of binding p53 in the presence of Nutlin concentrations that inhibit the wild-type HDM2-p53 interaction. The in vitro phenotypes were recapitulated in ex vivo assays measuring both p53 transactivation function and the direct p53-HDM2 interaction in the presence of Nutlin. Mutations conferring drug resistance were not confined to the N-terminal p53/Nutlin–binding domain, and were additionally seen in the acidic, zinc finger and RING domains. Mechanistic insights gleaned from this broad spectrum of mutations will aid in future drug design and further our understanding of the complex p53-HDM2 interaction. Published version 2014-01-21T06:23:49Z 2019-12-06T20:17:30Z 2014-01-21T06:23:49Z 2019-12-06T20:17:30Z 2013 2013 Journal Article Wei, S. J., Joseph, T., Sim, A. Y. L., Yurlova, L., Zolghadr, K., Lane, D., et al. (2013). In vitro selection of mutant HDM2 resistant to nutlin inhibition. PLoS One, 8(4), e62564-. https://hdl.handle.net/10356/100150 http://hdl.handle.net/10220/18644 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0062564 en PLoS one © 2013 The Authors. This paper was published in PLoS One and is made available as an electronic reprint (preprint) with permission of the authors. The paper can be found at the following official OpenURL: [http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0062564]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences
spellingShingle DRNTU::Science::Biological sciences
Wei, Siau Jia.
Joseph, Thomas.
Sim, Adelene Y. L.
Yurlova, Larisa.
Zolghadr, Kourosh.
Lane, David.
Verma, Chandra.
Ghadessy, Farid.
In vitro selection of mutant HDM2 resistant to nutlin inhibition
description HDM2 binds to the p53 tumour suppressor and targets it for proteosomal degradation. Presently in clinical trials, the small molecule Nutlin-3A competitively binds to HDM2 and abrogates its repressive function. Using a novel in vitro selection methodology, we simulated the emergence of resistance by evolving HDM2 mutants capable of binding p53 in the presence of Nutlin concentrations that inhibit the wild-type HDM2-p53 interaction. The in vitro phenotypes were recapitulated in ex vivo assays measuring both p53 transactivation function and the direct p53-HDM2 interaction in the presence of Nutlin. Mutations conferring drug resistance were not confined to the N-terminal p53/Nutlin–binding domain, and were additionally seen in the acidic, zinc finger and RING domains. Mechanistic insights gleaned from this broad spectrum of mutations will aid in future drug design and further our understanding of the complex p53-HDM2 interaction.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Wei, Siau Jia.
Joseph, Thomas.
Sim, Adelene Y. L.
Yurlova, Larisa.
Zolghadr, Kourosh.
Lane, David.
Verma, Chandra.
Ghadessy, Farid.
format Article
author Wei, Siau Jia.
Joseph, Thomas.
Sim, Adelene Y. L.
Yurlova, Larisa.
Zolghadr, Kourosh.
Lane, David.
Verma, Chandra.
Ghadessy, Farid.
author_sort Wei, Siau Jia.
title In vitro selection of mutant HDM2 resistant to nutlin inhibition
title_short In vitro selection of mutant HDM2 resistant to nutlin inhibition
title_full In vitro selection of mutant HDM2 resistant to nutlin inhibition
title_fullStr In vitro selection of mutant HDM2 resistant to nutlin inhibition
title_full_unstemmed In vitro selection of mutant HDM2 resistant to nutlin inhibition
title_sort in vitro selection of mutant hdm2 resistant to nutlin inhibition
publishDate 2014
url https://hdl.handle.net/10356/100150
http://hdl.handle.net/10220/18644
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0062564
_version_ 1759854235270250496