title: Stepwise assembly of functional C-terminal REST/NRSF transcriptional repressor complexes as a drug target.
abstract: In human cells, thousands of predominantly neuronal genes are regulated by the repressor element 1 (RE1)-silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF). REST/NRSF represses transcription of these genes in stem cells and non-neuronal cells by tethering corepressor complexes. Aberrant REST/NRSF expression and intracellular localization are associated with cancer and neurodegeneration in humans. To date, detailed molecular analyses of REST/NRSF and its C-terminal repressor complex have been hampered largely by the lack of sufficient amounts of purified REST/NRSF and its complexes. Therefore, the aim of this study was to express and purify human REST/NRSF and its C-terminal interactors in a baculovirus multiprotein expression system as individual proteins and coexpressed complexes. All proteins were enriched in the nucleus, and REST/NRSF was isolated as a slower migrating form, characteristic of nuclear REST/NRSF in mammalian cells. Both REST/NRSF alone and its C-terminal repressor complex were functionally active in histone deacetylation and histone demethylation and bound to RE1/neuron-restrictive silencer element (NRSE) sites. Additionally, the mechanisms of inhibition of the small-molecule drugs 4SC-202 and SP2509 were analyzed. These drugs interfered with the viability of medulloblastoma cells, where REST/NRSF has been implicated in cancer pathogenesis. Thus, a resource for molecular REST/NRSF studies and drug development has been established.
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Ken
authors.initials: K
authors.lastname: Inui
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Zongpei
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Juan
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Sakthidasan
authors.initials: S
authors.lastname: Jayaprakash
authors.affiliation: Centre for Stochastic Geometry and Advanced Bioimaging, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Le T M
authors.initials: LTM
authors.lastname: Le
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Srdja
authors.initials: S
authors.lastname: Drakulic
authors.affiliation: Centre for Stochastic Geometry and Advanced Bioimaging, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Bjoern
authors.initials: B
authors.lastname: Sander
authors.affiliation: Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark.|Institute of Human Genetics, Hannover Medical School, D-30625 Hannover, Germany.
authors.forename: Monika M
authors.initials: MM
authors.lastname: Golas
mesh_terms: D000107:Acetylation; D000818:Animals; D001549:Benzamides / Q000737:chemistry / Q000494:pharmacology; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006834:Hydrazines / Q000737:chemistry / Q000494:pharmacology; D046912:Multiprotein Complexes / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000072417:Protein Domains; D012097:Repressor Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology; D000099229:RE1-Silencing Transcription Factor
chemical_list: D001549:Benzamides; D006657:Histones; D006834:Hydrazines; D046912:Multiprotein Complexes; D009363:Neoplasm Proteins; D012097:Repressor Proteins; D013449:Sulfonamides; D000099229:RE1-Silencing Transcription Factor; C000594309:SP2509; C000614036:domatinostat
keywords: CoREST; DNA transcription; HDAC1; LSD1; REST/NRSF; drug mechanism; histone deacetylation; histone demethylation; medulloblastoma; transcriptional repression