The transcriptional repressor REST is a critical regulator of the neurosecretory phenotype

Authors

  • Alexander W. Bruce,

    1. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
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      The present address of Alexander W. Bruce is The Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, UK.

  • Alena Krejčí,

    1. Czech Academy of Sciences, Institute of Physiology, Prague, Czech Republic
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      The present address of Alena Krejčí is the Department of Anatomy, University of Cambridge, Cambridge CB2 1TN, UK.

  • Lezanne Ooi,

    1. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
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  • James Deuchars,

    1. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
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  • Ian C. Wood,

    1. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
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  • Vladimír Doležal,

    1. Czech Academy of Sciences, Institute of Physiology, Prague, Czech Republic
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  • Noel J. Buckley

    1. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK
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  • Note that we use the term gene silencer to describe a factor that completely blocks the synthesis of any target gene transcript to distinguish it from a repressor that results in lower numbers of detectable transcripts but nevertheless still some.

Address correspondence and reprint requests to Alexander Bruce, Transcription Regulation Group, The Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire CB10 1SA, UK.
E-mail: awb@sanger.ac.uk

Abstract

Release of distinct cellular cargoes in response to specific stimuli is a process fundamental to all higher eukaryotes and controlled by the regulated secretory pathway (RSP). However, the mechanism by which genes involved in the RSP are selectively expressed, leading to the establishment and appropriate functioning of regulated secretion remaining largely unknown. Using the rat pheochromocytoma cell line PC12, we provide evidence that, by controlling expression of many genes involved in the RSP, the transcriptional repressor REST can regulate this pathway and hence the neurosecretory phenotype. Introduction of REST transgenes into PC12 cells leads to the repression of many genes, the products of which are involved in regulated secretion. Moreover, chromatin immunoprecipitation assays show that many of the repressed genes recruit the recombinant REST protein to RE1 sites within their promoters and abrogation of REST function leads to reactivation of these transcripts. In addition to the observed transcriptional effects, PC12 cells expressing REST have fewer secretory granules and a reduction in the ability to store and release noradrenaline. Furthermore, an important trigger for synaptic release, influx of calcium through voltage-operated calcium channels, is compromised. This is the first demonstration of a transcription factor that directly controls expression of many major components of the RSP and provides further insight into the function of REST.

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