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Vol. 9, Issue 8, 2107-2123, August 1998
on Cell Cycle
Events
Department of Pathology, Stanford University School of Medicine,
Stanford, California 94305-5324
Pol
is the principal DNA polymerase for initiation of DNA
replication and also functions in postinitiation DNA synthesis. In this
study, we investigated the cell cycle responses induced by mutations in
pol+. Germinating spores carrying either
a deletion of pol+
(pol
) or a structurally intact but catalytically
dead pol mutation proceed to inappropriate mitosis
with no DNA synthesis. This suggests that the catalytic function, and
not the physical presence of Pol, is required to generate the signal
that prevents the cells from entering mitosis prematurely. Cells with a
polts allele arrest the cell cycle near the
hydroxyurea arrest point, but, surprisingly, polts in
cdc20 (pol
mutant) background arrested with a cdc
phenoytpe, not a polts-like phenotype. At 25°C,
replication perturbation caused by polts alleles
induces Cds1 kinase activity and requires the checkpoint Rads, Cds1,
and Rqh1, but not Chk1, to maintain cell viability. At 36°C,
replication disruption caused by polts alleles
induces the phosphorylation of Chk1; however, mutant cells arrest with
heterogeneous cell sizes with a population of the cells entering
aberrant mitosis. Together, our results indicate that the initiation
DNA structure synthesized by Pol is required to bring about the S
phase to mitosis checkpoint, whereas replication defects of different
severity caused by polts mutations induce
differential downstream kinase responses.
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