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An ACF1–ISWI chromatin-remodeling complex is required for DNA replication through heterochromatin

Nature Genetics volume 32, pages 627–632 (2002)Cite this article

Abstract

The mechanism by which the eukaryotic DNA-replication machinery penetrates condensed chromatin structures to replicate the underlying DNA is poorly understood. Here we provide evidence that an ACF1–ISWI chromatin-remodeling complex is required for replication through heterochromatin in mammalian cells. ACF1 (ATP-utilizing chromatin assembly and remodeling factor 1) and an ISWI isoform, SNF2H (sucrose nonfermenting-2 homolog), become specifically enriched in replicating pericentromeric heterochromatin. RNAi-mediated depletion of ACF1 specifically impairs the replication of pericentromeric heterochromatin. Accordingly, depletion of ACF1 causes a delay in cell-cycle progression through the late stages of S phase. In vivo depletion of SNF2H slows the progression of DNA replication throughout S phase, indicating a functional overlap with ACF1. Decondensing the heterochromatin with 5-aza-2-deoxycytidine reverses the effects of ACF1 and SNF2H depletion. Expression of an ACF1 mutant that cannot interact with SNF2H also interferes with replication of condensed chromatin. Our data suggest that an ACF1–SNF2H complex is part of a dedicated mechanism that enables DNA replication through highly condensed regions of chromatin.

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Figure 1: ACF1 and SNF2H target replicating pericentromeric heterochromatin.
Figure 2: Depletion of ACF1 by RNAi interferes with late S-phase DNA replication.
Figure 3: 5-aza-2-deoxycytidine abolishes the requirement of ACF1 for DNA replication.
Figure 4: Depletion of SNF2H by RNAi impedes DNA replication throughout S phase.
Figure 5: ACF1–SNF2H interaction is important for ACF1 function in vivo.

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Acknowledgements

We thank Y. Nakamura for hSNF2H cDNA; L. Bozhenok, M. Grimaldi, G. Elliot, R. Cross, J. Dalgaard, W. Bickmore and J. Boyes for helpful discussions and assistance in various aspects of this project; G. Almouzni and M. Guenatri for advice and discussion; and G. Almouzni for hosting R.A.P. under a network grant from the European Union. This work was funded by the Marie Curie Cancer Care. R.A.P. was supported by a grant from the Association for International Cancer Research, St Andrews. I.K. was supported in part by a research fellowship from the Uehara Memorial Foundation.

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Author notes

  1. Graham Dellaire

    Present address: Cell Biology Program, The Hospital for Sick Children Research Institute, Toronto, Canada

  2. Nadine Collins and Raymond A. Poot: These authors contributed equally to this work.

Authors and Affiliations

  1. Marie Curie Research Institute, The Chart, Oxted, Surrey, RH8 0TL, UK

    Nadine Collins, Raymond A. Poot, Iwao Kukimoto, Custodia García-Jiménez & Patrick D. Varga-Weisz

  2. Medical Research Council Human Genetics Unit, Edinburgh, UK

    Graham Dellaire

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Collins, N., Poot, R., Kukimoto, I. et al. An ACF1–ISWI chromatin-remodeling complex is required for DNA replication through heterochromatin. Nat Genet 32, 627–632 (2002). https://doi.org/10.1038/ng1046

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