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General transcription factors bind promoters repressed by Polycomb group proteins

Nature volume 412, pages 651–655 (2001)Cite this article

Abstract

To maintain cell identity during development and differentiation, mechanisms of cellular memory have evolved that preserve transcription patterns in an epigenetic manner. The proteins of the Polycomb group (PcG) are part of such a mechanism, maintaining gene silencing. They act as repressive multiprotein complexes that may render target genes inaccessible to the transcriptional machinery1,2, inhibit chromatin remodelling3,4, influence chromosome domain topology5 and recruit histone deacetylases (HDACs)6. PcG proteins have also been found to bind to core promoter regions7, but the mechanism by which they regulate transcription remains unknown. To address this, we used formaldehyde-crosslinked chromatin immunoprecipitation (X-ChIP) to map TATA-binding protein (TBP), transcription initiation factor IIB (TFIIB) and IIF (TFIIF), and dHDAC1 (RPD3) across several Drosophila promoter regions. Here we show that binding of PcG proteins to repressed promoters does not exclude general transcription factors (GTFs) and that depletion of PcG proteins by double-stranded RNA interference leads to de-repression of developmentally regulated genes. We further show that PcG proteins interact in vitro with GTFs. We suggest that PcG complexes maintain silencing by inhibiting GTF-mediated activation of transcription.

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Figure 1: Analysis of the gene expression and acetylation state of BX-C, engrailed, empty spiracles, RpII140 and brown genes.
Figure 2: PCR analysis of the protein distribution at BX-C, engrailed, empty spiracles, RpII140 and brown promoter regions.
Figure 3: Interaction of TBP with PcG proteins.
Figure 4: Post-transcriptional silencing of Pc and ph in SL-2 cells.

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Acknowledgements

We thank B. M. Paterson for advice concerning the RNAi experiments and J. Kadonaga for discussions and comments on the manuscript. We are indebted to J. Butler, J. Kadonaga, A. Barlow and R. Paro for providing (respectively) dTBP, dTFIIB, dTFIIF, dHDAC1 and PC antibodies. We thank N. Collu for technical assistance. This work was supported by a postdoctoral fellowship to A.B. by EU-TMR, and by research grants from A.I.R.C. and TELETHON to V.O., from the Wellcome Trust and the Human Frontier Science Program to B.M.T. and from MURST to M.E.B.

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Authors and Affiliations

  1. DIBIT, San Raffaele Scientific Institute, Via Olgettina 58, Milan, 20132, Italy

    Achim Breiling, Marco E. Bianchi & Valerio Orlando

  2. Chromatin and Gene Expression Group, University of Birmingham Medical School, Birmingham, B15 2TT, UK

    Bryan M. Turner

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  1. Achim Breiling
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Correspondence to Valerio Orlando.

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Breiling, A., Turner, B., Bianchi, M. et al. General transcription factors bind promoters repressed by Polycomb group proteins. Nature 412, 651–655 (2001). https://doi.org/10.1038/35088090

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