Abstract
MCM10 is essential for the initiation of chromosomal DNA replication in Saccharomyces cerevisiae. Previous work showed that Mcm10p interacts with the Mcm2–7 protein complex that may be functioning as the replication‐licensing factor. In addition, Mcm10p is required during origin activation and disassembly of the prereplicative complex, which allows smooth passage of replication forks.
We show that an mcm10 mutation causes a slow progression of DNA synthesis and a loss of chromosome integrity during the S phase and prevents entry into mitosis, despite apparent completion of chromosomal DNA replication at nonpermissive temperatures. Furthermore, Mcm10p interacts genetically with the origin recognition complex (ORC) and various replication elongation factors, including a subunit of DNA polymerases ε and δ. Mcm10p is an abundant protein (approximately 4 × 104 copies per haploid cell) that is almost exclusively localized in the chromatin and/or nuclear matrix fractions during all phases of the cell cycle. When it is visualized by the chromosome‐spreading method followed by immunostaining, Mcm10p forms punctate foci on chromatin throughout the cell cycle and these foci mostly overlap with those of Orc1p, a component of ORC.
These results suggest that Mcm10p, like the Mcm2–7 proteins, is a critical component of the prereplication chromatin and acts together with ORC during the initiation of chromosomal DNA replication; in addition, Mcm10p plays an important role during the elongation of DNA replication.
We show that an mcm10 mutation causes a slow progression of DNA synthesis and a loss of chromosome integrity during the S phase and prevents entry into mitosis, despite apparent completion of chromosomal DNA replication at nonpermissive temperatures. Furthermore, Mcm10p interacts genetically with the origin recognition complex (ORC) and various replication elongation factors, including a subunit of DNA polymerases ε and δ. Mcm10p is an abundant protein (approximately 4 × 104 copies per haploid cell) that is almost exclusively localized in the chromatin and/or nuclear matrix fractions during all phases of the cell cycle. When it is visualized by the chromosome‐spreading method followed by immunostaining, Mcm10p forms punctate foci on chromatin throughout the cell cycle and these foci mostly overlap with those of Orc1p, a component of ORC.
These results suggest that Mcm10p, like the Mcm2–7 proteins, is a critical component of the prereplication chromatin and acts together with ORC during the initiation of chromosomal DNA replication; in addition, Mcm10p plays an important role during the elongation of DNA replication.
| Original language | English |
|---|---|
| Pages (from-to) | 975-989 |
| Number of pages | 15 |
| Journal | Genes to Cells |
| Volume | 5 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - Dec 2000 |
| Externally published | Yes |
Bibliographical note
AcknowledgementsWe thank Drs S. P. Bell, O. M. Aparicio, T. Tanaka, J. Rine, K. Shirahige, H. Araki, Y. Kamimura, N. Nakashima and H. Masumoto for providing strains, plasmids, and antibody. We also thank Dr B. K. Tye for discussion during the course of this study. This work was supported partly by a Grant‐in‐Aid for Scientific Research on Priority Area (A) and (B) from the Ministry of Education, Science, Sports, and Culture of Japan to A.S. and Y.K., respectively