Stepwise assembly of functional C-terminal REST/NRSF transcriptional repressor complexes as a drug target

作者：Inui Ken; Zhao Zongpei; Yuan Juan; Jayaprakash Sakthidasan; Le Le T M; Drakulic Srdja; Sander Bjoern; Golas Monika M

来源：Protein Science, 2017, 26(5): 997-1011.

DOI：10.1002/pro.3142

• REST
• NRSF
• CoREST
• LSD1
• HDAC1
• DNA transcription
• transcriptional repression
• histone deacetylation
• histone demethylation
• drug mechanism
• medulloblastoma

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.

• 出版日期2017-5