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Vol. 10, Issue 10, 3331-3343, October 1999
and
Department of Biochemistry and Molecular Biology, The University of
Tokyo, Graduate School of Medicine, Tokyo 113-0033, Japan.
Fission yeast rad22+, a homologue of
budding yeast RAD52, encodes a double-strand break
repair component, which is dispensable for proliferation. We, however,
have recently obtained a cell division cycle mutant with a
temperature-sensitive allele of rad22+,
designated rad22-H6, which resulted from a point
mutation in the conserved coding sequence leading to one amino acid
alteration. We have subsequently isolated
rad22+ and its novel homologue
rti1+ as multicopy suppressors of this
mutant. rti1+ suppresses all the defects of
cells lacking rad22+. Mating type
switch-inactive heterothallic cells lacking either rad22+ or rti1+
are viable, but those lacking both genes are inviable and arrest proliferation with a cell division cycle phenotype. At the
nonpermissive temperature, a synchronous culture of
rad22-H6 cells performs DNA synthesis without delay and
arrests with chromosomes seemingly intact and replication completed and
with a high level of tyrosine-phosphorylated Cdc2. However,
rad22-H6 cells show a typical S phase arrest phenotype if combined with the rad1-1 checkpoint mutation.
rad22+ genetically interacts with
rad11+, which encodes the large subunit of
replication protein A. Deletion of
rad22+/rti1+ or
the presence of rad22-H6 mutation decreases the
restriction temperature of rad11-A1 cells by 4-6°C
and leads to cell cycle arrest with chromosomes incompletely
replicated. Thus, in fission yeast a double-strand break repair
component is required for a certain step of chromosome replication
unlinked to repair, partly via interacting with replication protein A.
Department of Bioengineering, Faculty
of Engineering, Soka University, Tangi-cho, Hachioji, Tokyo 192, Japan;
and
Cell Cycle Laboratory, Imperial Cancer Research
Fund, Lincoln's Inn Fields, London WC2A 3PX, UK.
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