Expression of the Neural REST/NRSF–SIN3 Transcriptional Corepressor Complex as a Target for Small-Molecule Inhibitors

Sakthidasan Jayaprakash; Le T.M. Le; Bjoern Sander; Monika M. Golas

doi:10.1007/s12033-020-00283-7

Expression of the Neural REST/NRSF–SIN3 Transcriptional Corepressor Complex as a Target for Small-Molecule Inhibitors

Sakthidasan Jayaprakash, Le T.M. Le, Bjoern Sander, Monika M. Golas^*

^*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

Abstract

The repressor element 1 (RE1) silencing transcription factor/neuron-restrictive silencing factor (REST/NRSF) modulates the expression of genes with RE1/neuron-restrictive silencing element (RE1/NRSE) sites by recruiting the switch independent 3 (SIN3) factor and the REST corepressor (COREST) to its N and C-terminal repressor domain, respectively. Both, SIN3 and COREST assemble into protein complexes that are composed of multiple subunits including a druggable histone deacetylase (HDAC) enzyme. The SIN3 core complex comprises the eponymous proteins SIN3A or SIN3B, the catalytically active proteins HDAC1 or HDAC2, the histone chaperone retinoblastoma-associated protein 46/retinoblastoma-binding protein 7 (RBAP46/RBBP7) or RBAP48/RBBP4, the SIN3-associated protein 30 (SAP30), and the suppressor of defective silencing 3 (SDS3). Here, we overcome a bottleneck limiting the molecular characterization of the REST/NRSF–SIN3 transcriptional corepressor complex. To this end, SIN3 genes were amplified from the complementary DNA of neural stem/progenitor cells, and expressed in a baculovirus/insect cell expression system. We show that the isolates bind to DNA harboring RE1/NRSE sites and demonstrate that the histone deacetylase activity is blocked by small-molecule inhibitors. Thus, our isolates open up for future biomedical research on this critical transcriptional repressor complex and are envisioned as tool for drug testing.

Originalsprog	Engelsk
Tidsskrift	Molecular Biotechnology
Vol/bind	63
Nummer	1
Sider (fra-til)	53-62
Antal sider	10
ISSN	1073-6085
DOI	https://doi.org/10.1007/s12033-020-00283-7
Status	Udgivet - jan. 2021

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title = "Expression of the Neural REST/NRSF–SIN3 Transcriptional Corepressor Complex as a Target for Small-Molecule Inhibitors",

abstract = "The repressor element 1 (RE1) silencing transcription factor/neuron-restrictive silencing factor (REST/NRSF) modulates the expression of genes with RE1/neuron-restrictive silencing element (RE1/NRSE) sites by recruiting the switch independent 3 (SIN3) factor and the REST corepressor (COREST) to its N and C-terminal repressor domain, respectively. Both, SIN3 and COREST assemble into protein complexes that are composed of multiple subunits including a druggable histone deacetylase (HDAC) enzyme. The SIN3 core complex comprises the eponymous proteins SIN3A or SIN3B, the catalytically active proteins HDAC1 or HDAC2, the histone chaperone retinoblastoma-associated protein 46/retinoblastoma-binding protein 7 (RBAP46/RBBP7) or RBAP48/RBBP4, the SIN3-associated protein 30 (SAP30), and the suppressor of defective silencing 3 (SDS3). Here, we overcome a bottleneck limiting the molecular characterization of the REST/NRSF–SIN3 transcriptional corepressor complex. To this end, SIN3 genes were amplified from the complementary DNA of neural stem/progenitor cells, and expressed in a baculovirus/insect cell expression system. We show that the isolates bind to DNA harboring RE1/NRSE sites and demonstrate that the histone deacetylase activity is blocked by small-molecule inhibitors. Thus, our isolates open up for future biomedical research on this critical transcriptional repressor complex and are envisioned as tool for drug testing.",

keywords = "HDAC, REST/NRSF, SIN3, Small-molecule inhibitors, Transcriptional repression",

author = "Sakthidasan Jayaprakash and Le, {Le T.M.} and Bjoern Sander and Golas, {Monika M.}",

note = "Funding Information: We are grateful to Golshah Ayoubi and Susanne N. Stubbe for excellent technical assistance, and wish to thank Zongpei Zhao for technical support. We acknowledge access to laboratory facilities at the Danish Neuroscience Center House. This work has been supported by the Lundbeck Foundation{\textquoteright}s Fellowship program, the Sapere Aude Program of the Danish Council for Independent Research, the Danish Cancer Society, the Carlsberg Foundation, the A.P. M{\o}ller Foundation for the Advancement of Medical Sciences, the Fabrikant Einar Willumsens Mindelegat and the Helga og Peter Kornings Fond to M.M.G. Publisher Copyright: {\textcopyright} 2020, Springer Science+Business Media, LLC, part of Springer Nature. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

year = "2021",

month = jan,

doi = "10.1007/s12033-020-00283-7",

language = "English",

volume = "63",

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Expression of the Neural REST/NRSF–SIN3 Transcriptional Corepressor Complex as a Target for Small-Molecule Inhibitors. / Jayaprakash, Sakthidasan; Le, Le T.M.; Sander, Bjoern et al.
I: Molecular Biotechnology, Bind 63, Nr. 1, 01.2021, s. 53-62.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

TY - JOUR

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AU - Jayaprakash, Sakthidasan

AU - Le, Le T.M.

AU - Sander, Bjoern

AU - Golas, Monika M.

N1 - Funding Information: We are grateful to Golshah Ayoubi and Susanne N. Stubbe for excellent technical assistance, and wish to thank Zongpei Zhao for technical support. We acknowledge access to laboratory facilities at the Danish Neuroscience Center House. This work has been supported by the Lundbeck Foundation’s Fellowship program, the Sapere Aude Program of the Danish Council for Independent Research, the Danish Cancer Society, the Carlsberg Foundation, the A.P. Møller Foundation for the Advancement of Medical Sciences, the Fabrikant Einar Willumsens Mindelegat and the Helga og Peter Kornings Fond to M.M.G. Publisher Copyright: © 2020, Springer Science+Business Media, LLC, part of Springer Nature. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/1

Y1 - 2021/1

N2 - The repressor element 1 (RE1) silencing transcription factor/neuron-restrictive silencing factor (REST/NRSF) modulates the expression of genes with RE1/neuron-restrictive silencing element (RE1/NRSE) sites by recruiting the switch independent 3 (SIN3) factor and the REST corepressor (COREST) to its N and C-terminal repressor domain, respectively. Both, SIN3 and COREST assemble into protein complexes that are composed of multiple subunits including a druggable histone deacetylase (HDAC) enzyme. The SIN3 core complex comprises the eponymous proteins SIN3A or SIN3B, the catalytically active proteins HDAC1 or HDAC2, the histone chaperone retinoblastoma-associated protein 46/retinoblastoma-binding protein 7 (RBAP46/RBBP7) or RBAP48/RBBP4, the SIN3-associated protein 30 (SAP30), and the suppressor of defective silencing 3 (SDS3). Here, we overcome a bottleneck limiting the molecular characterization of the REST/NRSF–SIN3 transcriptional corepressor complex. To this end, SIN3 genes were amplified from the complementary DNA of neural stem/progenitor cells, and expressed in a baculovirus/insect cell expression system. We show that the isolates bind to DNA harboring RE1/NRSE sites and demonstrate that the histone deacetylase activity is blocked by small-molecule inhibitors. Thus, our isolates open up for future biomedical research on this critical transcriptional repressor complex and are envisioned as tool for drug testing.

AB - The repressor element 1 (RE1) silencing transcription factor/neuron-restrictive silencing factor (REST/NRSF) modulates the expression of genes with RE1/neuron-restrictive silencing element (RE1/NRSE) sites by recruiting the switch independent 3 (SIN3) factor and the REST corepressor (COREST) to its N and C-terminal repressor domain, respectively. Both, SIN3 and COREST assemble into protein complexes that are composed of multiple subunits including a druggable histone deacetylase (HDAC) enzyme. The SIN3 core complex comprises the eponymous proteins SIN3A or SIN3B, the catalytically active proteins HDAC1 or HDAC2, the histone chaperone retinoblastoma-associated protein 46/retinoblastoma-binding protein 7 (RBAP46/RBBP7) or RBAP48/RBBP4, the SIN3-associated protein 30 (SAP30), and the suppressor of defective silencing 3 (SDS3). Here, we overcome a bottleneck limiting the molecular characterization of the REST/NRSF–SIN3 transcriptional corepressor complex. To this end, SIN3 genes were amplified from the complementary DNA of neural stem/progenitor cells, and expressed in a baculovirus/insect cell expression system. We show that the isolates bind to DNA harboring RE1/NRSE sites and demonstrate that the histone deacetylase activity is blocked by small-molecule inhibitors. Thus, our isolates open up for future biomedical research on this critical transcriptional repressor complex and are envisioned as tool for drug testing.

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DO - 10.1007/s12033-020-00283-7

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Expression of the Neural REST/NRSF–SIN3 Transcriptional Corepressor Complex as a Target for Small-Molecule Inhibitors

Abstract

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