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Nucleic Acids Research, 1990, Vol. 18, No. 19 5767-5774
© 1990


MOLECULAR BIOLOGY

Chromatin assembly on replicating DNA in vitro

Geneviéve Almouzni1,2, David J. Clark1, Marcel Méchali2 and Alan P. Wolffe1,*

1Laboratory of Molecular Biology, NIDDK, National Institutes of Health Bethesda, MD 20892, USA 2Institut Jacques Monod, CNRS, Université Paris 7, Tour 43, 2 Place Jussieu F-75251, Paris Cedex 05, France

*To whom correspondence should be addressed

Received June 13, 1990. Revised August 21, 1990. Accepted August 21, 1990.

Replicating single-stranded DNA is preferentially assembled Into chromatin in Xenopus egg extracts relative to non-replicating double-stranded DNA. We have examined the molecular basis of this phenomenon. Single-stranded DNA itself is not a favored template for nucleosome assembly in comparison to double-stranded DNA. Complementary strand synthesis is required for the rapid assembly of nucleosomes. We present evidence that the assembly of chromatin on replicating DNA is a two step phenomenon. The first step involves the replication of DNA and the assembly of an intermediate structure, the second step involves the sequestration of histories H2A/H2B onto DNA. Histones H2A/H2B are preferentially sequestered onto replicated DNA in comparison to non-replicated DNA incubated in the extract.


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