Abstract
The assembly of DNA into chromatin is a critical step in the replication and repair of the eukaryotic genome1,2,3,4,5,6,7,8. It has been known for nearly 20 years that chromatin assembly is an ATP-dependent process9. ATP-dependent chromatin-assembly factor (ACF) uses the energy of ATP hydrolysis for the deposition of histones into periodic nucleosome arrays, and the ISWI subunit of ACF is an ATPase that is related to helicases10,11. Here we show that ACF becomes committed to the DNA template upon initiation of chromatin assembly. We also observed that ACF assembles nucleosomes in localized arrays, rather than randomly distributing them. By using a purified ACF-dependent system for chromatin assembly, we found that ACF hydrolyses about 2–4 molecules of ATP per base pair in the assembly of nucleosomes. This level of ATP hydrolysis is similar to that used by DNA helicases for the unwinding of DNA12. These results suggest that a tracking mechanism exists in which ACF assembles chromatin as an ATP-driven DNA-translocating motor. Moreover, this proposed mechanism for ACF may be relevant to the function of other chromatin-remodelling factors that contain ISWI subunits.
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Acknowledgements
We thank P. Geiduschek, R. Dutnall, L. Pillus, T. Juven-Gershon, V. Alexiadis, T. Boulay, B. Santoso, A. Lusser, J. Huang Parsons and M. Levenstein for critical reading of the manuscript. This work was supported by a grant from the National Institutes of Health to J.T.K. D.V.F. is an American Cancer Society Postdoctoral Fellow.
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Fyodorov, D., Kadonaga, J. Dynamics of ATP-dependent chromatin assembly by ACF. Nature 418, 896–900 (2002). https://doi.org/10.1038/nature00929
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DOI: https://doi.org/10.1038/nature00929
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