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

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DOI

  • Sakthidasan Jayaprakash
  • ,
  • Le T.M. Le
  • ,
  • Bjoern Sander
  • Monika M. Golas, Hannover Medical School

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.

OriginalsprogEngelsk
TidsskriftMolecular Biotechnology
Vol/bind63
Nummer1
Sider (fra-til)53-62
Antal sider10
ISSN1073-6085
DOI
StatusUdgivet - jan. 2021

Bibliografisk 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’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.

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