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Vol. 14, No. 8, pp. 913-926, April 15, 2000
Department of Molecular Biology and Genetics, Cornell University,
Ithaca, New York 14853-2703 USA
MCM2-7, a complex of six subunits, is an essential component of the
prereplication chromatin that is assembled at Saccharomyces cerevisiae replication origins during G1 phase. It is
also believed to be the processive helicase at growing forks. To
elucidate the action of MCM2-7 during the transition from initiation
to elongation replication, we have focused our studies on Mcm10, a
replication initiation protein that physically interacts with members
of the MCM2-7 complex. We show that Mcm10 is a chromatin-associated
protein that mediates the association of the MCM2-7 complex with
replication origins. Furthermore, diminished interaction between Mcm10
and Mcm7, a subunit of the MCM2-7 complex, by a mutation in either Mcm10 or Mcm7 inhibits replication initiation. Surprisingly, a double
mutant containing both the mcm10-1 and mcm7-1
(cdc47-1) alleles restores interaction between Mcm10 and Mcm7
and corrects all of the defects exhibited by each of the single
mutants, including the stalling of replication forks at replication
origins typically seen in mcm10-1 cells. This mutual
compensation of defects between two independently isolated mutations is
allele specific. These results suggest that Mcm10, like Mcm7, is a
critical component of the prereplication chromatin and that interaction
between Mcm10 and Mcm7 is required for proper replication initiation
and prompt release of origin-bound factors.
[Key Words: MCM2-7 complex; Mcm10; Cdc45; DNA synthesis; replication initiation]
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