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Vol. 12, Issue 11, 3658-3667, November 2001
and
*ICRF Clare Hall Laboratories, South Mimms, Hertfordshire, EN6 3LD,
United Kingdom; A prereplicative complex (pre-RC) of proteins is assembled at
budding yeast origins of DNA replication during the G1-phase of the
cell cycle, as shown by genomic footprinting. The proteins responsible
for this prereplicative footprint have yet to be identified but are
likely to be involved in the earliest stages of the initiation step of
chromosome replication. Here we show that MCM2-7 proteins are
essential for both the formation and maintenance of the pre-RC footprint at the origin ARS305. It is likely that pre-RCs contain heteromeric complexes of MCM2-7 proteins, since degradation of Mcm2,
3, 6, or 7 during G1-phase, after pre-RC formation, causes loss of Mcm4
from the nucleus. It has been suggested that pre-RCs on unreplicated
chromatin may generate a checkpoint signal that inhibits premature
mitosis during S-phase. We show that, although mitosis does indeed
occur in the absence of replication if MCM proteins are degraded during
G1-phase, anaphase is prevented if MCMs are degraded during S-phase.
Our data indicate that pre-RCs do not play a direct role in checkpoint
control during chromosome replication.
Department of Zoology, University of
Oxford, Oxford, OX1 3PS, United Kingdom
Corresponding author. E-mail address:
J.Diffley{at}icrf.icnet.uk.
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