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Expression of the Neural REST/NRSF-SIN3 Transcriptional Corepressor Complex as a Target for Small-Molecule Inhibitors.
- Source :
-
Molecular biotechnology [Mol Biotechnol] 2021 Jan; Vol. 63 (1), pp. 53-62. Date of Electronic Publication: 2020 Nov 01. - Publication Year :
- 2021
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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.
- Subjects :
- Animals
Baculoviridae metabolism
Benzamides pharmacology
Co-Repressor Proteins isolation & purification
Co-Repressor Proteins metabolism
Depsipeptides pharmacology
Gene Library
Histone Deacetylases metabolism
Humans
Nerve Tissue Proteins isolation & purification
Nerve Tissue Proteins metabolism
Neural Stem Cells enzymology
Pyrimidines pharmacology
Recombinant Proteins
Regulatory Elements, Transcriptional genetics
Repressor Proteins isolation & purification
Repressor Proteins metabolism
Sf9 Cells
Sin3 Histone Deacetylase and Corepressor Complex metabolism
Co-Repressor Proteins genetics
Histone Deacetylase Inhibitors pharmacology
Nerve Tissue Proteins genetics
Neural Stem Cells metabolism
Repressor Proteins genetics
Sin3 Histone Deacetylase and Corepressor Complex genetics
Sin3 Histone Deacetylase and Corepressor Complex isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1559-0305
- Volume :
- 63
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Molecular biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 33130996
- Full Text :
- https://doi.org/10.1007/s12033-020-00283-7