Nucleic Acids Research, Vol 27, Issue 10 2175-2180, Copyright © 1999 by Oxford University Press
K Ohta, TC Wu, M Lichten and T Shibata
In Saccharomyces cerevisiae, DNA double-strand breaks (DSBs) initiate
meiotic recombination at open sites in chromatin, which display a
meiosis-specific increase in micrococcal nuclease (MNase) sensitivity. The
arg4 promoter contains such a DSB site. When arg4 sequences are placed in a
pBR322-derived insert at HIS4 (his4 :: arg4 ), the presence of strong DSB
sites in pBR322 sequences leads to an almost complete loss of breaks from
the insert-borne arg4 promoter region. Most of the MNase-sensitive sites
occurred at similar positions in insert-borne and in normal ARG4 sequences,
indicating that hotspot inactivation is not a consequence of changes in
nucleosome positioning. However, a meiosis- specific increase in MNase
hypersensitivity was no longer detected at the inactive insert-borne arg4
DSB site. Elimination of pBR322 sequences restored DSBs to the insert-borne
arg4 promoter region and also restored the meiotic induction of MNase
hypersensitivity. Thus, the meiotic induction of MNase hypersensitivity at
the DSB sites is suppressed and activated in parallel to DSBs themselves,
without changes in the underlying DNA sequence or nucleosome positioning.
We suggest that meiosis-specific changes in chromatin at a DSB site are a
signal reflecting a pivotal step in DSB formation.
ARTICLES
Competitive inactivation of a double-strand DNA break site involves parallel suppression of meiosis-induced changes in chromatin configuration
Cellular and Molecular Biology Laboratory, The Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama 351-0198, Japan. kohta@postman.riken.go.jp
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