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RNA interference demonstrates a novel role for H2A.Z in chromosome segregation

Nature Structural & Molecular Biology volume 11, pages 650–655 (2004)Cite this article

Abstract

The histone variant H2A.Z plays an essential role in metazoans but its function remains to be determined. Here, we developed a new inducible RNAi strategy to elucidate the role of H2A.Z in mammalian cell lines. We show that in the absence of H2A.Z, the genome becomes highly unstable and that this instability is caused by defects in the chromosome segregation process. Analysis of H2A.Z localization reveals that in these cells it is enriched at heterochromatic foci with HP1α on the arms of chromosomes but not at centromeric regions. When H2A.Z is depleted, normal HP1α-chromatin interactions are disrupted on the chromosomal arms and, notably, also at pericentric regions. Therefore, H2A.Z controls the localization of HP1α. We conclude that H2A.Z is essential for the accurate transmission of chromosomes.

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Figure 1: Inducible inhibition of H2A.Z expression.
Figure 2: Inhibition of H2A.Z expression produces an unstable genome.
Figure 3: Chromosome segregation defects in H2A.Z siRNA-treated cells.
Figure 4: HP1α localization to the arms of chromosomes is lost in H2A.Z siRNA-treated cells.
Figure 5: Disruption of proper HP1α-chromatin interactions in the absence of H2A.Z.

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Acknowledgements

This work was supported by a grant from the Australian National Health and Medical Research Council to D.T. We thank P. Ridgway for critical reading of the manuscript.

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Authors and Affiliations

  1. The John Curtin School of Medical Research, The Australian National University, PO Box 334, Canberra, Australian Capital Territory, 2601, Australia

    Danny Rangasamy, Ian Greaves & David J Tremethick

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  1. Danny Rangasamy
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  2. Ian Greaves
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  3. David J Tremethick
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Correspondence to David J Tremethick.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

H2A.Z expression can be inhibited by siRNA. (PDF 207 kb)

Supplementary Fig. 2

Inducible synthesis of H2A.Z siRNA. (PDF 151 kb)

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Rangasamy, D., Greaves, I. & Tremethick, D. RNA interference demonstrates a novel role for H2A.Z in chromosome segregation. Nat Struct Mol Biol 11, 650–655 (2004). https://doi.org/10.1038/nsmb786

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