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Role of histone H2A ubiquitination in Polycomb silencing

Nature volume 431, pages 873–878 (2004)Cite this article

Abstract

Covalent modification of histones is important in regulating chromatin dynamics and transcription1,2. One example of such modification is ubiquitination, which mainly occurs on histones H2A and H2B3. Although recent studies have uncovered the enzymes involved in histone H2B ubiquitination4,5,6 and a ‘cross-talk’ between H2B ubiquitination and histone methylation7,8, the responsible enzymes and the functions of H2A ubiquitination are unknown. Here we report the purification and functional characterization of an E3 ubiquitin ligase complex that is specific for histone H2A. The complex, termed hPRC1L (human Polycomb repressive complex 1-like), is composed of several Polycomb-group proteins including Ring1, Ring2, Bmi1 and HPH2. hPRC1L monoubiquitinates nucleosomal histone H2A at lysine 119. Reducing the expression of Ring2 results in a dramatic decrease in the level of ubiquitinated H2A in HeLa cells. Chromatin immunoprecipitation analysis demonstrated colocalization of dRing with ubiquitinated H2A at the PRE and promoter regions of the Drosophila Ubx gene in wing imaginal discs. Removal of dRing in SL2 tissue culture cells by RNA interference resulted in loss of H2A ubiquitination concomitant with derepression of Ubx. Thus, our studies identify the H2A ubiquitin ligase, and link H2A ubiquitination to Polycomb silencing.

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Figure 1: Identification of an H2A ubiquitin ligase activity in HeLa cells.
Figure 2: Purification and identification of the H2A ubiquitination ligase complex.
Figure 3: Ring2 is the catalytic subunit and is required for H2A ubiquitination in vivo.
Figure 4: dRing-dependent H2A ubiquitination at Ubx PRE and promoter regions.

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Acknowledgements

We thank J. Kim for help with mass spectrometry. This work was supported by NIH grants to Y.Z., R.S.J. and P.T.

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, 27599-7295, USA

    Hengbin Wang & Yi Zhang

  2. Department of Biological Sciences, Southern Methodist University, Dallas, Texas, 75275, USA

    Liangjun Wang & Richard S. Jones

  3. Molecular Biology Program, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, 10021, New York, USA

    Hediye Erdjument-Bromage & Paul Tempst

  4. Departmamento de Desarrollo y Biologia Celular, Centro de Investigaciones Biologicas, Ramiro de Maeztu, 9, E28040, Madrid, Spain

    Miguel Vidal

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  1. Hengbin Wang
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Correspondence to Yi Zhang.

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Supplementary information

Supplementary Data

The monoclonal uH2A antibody recognizes uH2A from Drosophila SL2 cells. (DOC 113 kb)

Supplementary Methods

Purification and identification of histone H2A ubiquitin ligase complex. 2. Generation and characterization of Ring2 knock-down cell lines. 3. Extraction of ubiquitinated histones from Drosophila SL2 cells. (DOC 53 kb)

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Wang, H., Wang, L., Erdjument-Bromage, H. et al. Role of histone H2A ubiquitination in Polycomb silencing. Nature 431, 873–878 (2004). https://doi.org/10.1038/nature02985

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