Abstract
Mcm10 is a conserved eukaryotic DNA replication factor whose function has remained elusive. We report here that Mcm10 binding to replication origins in budding yeast is cell cycle regulated and dependent on the putative helicase, Mcm2-7. Mcm10 is also an essential component of the replication fork. A fraction of Mcm10 binds to DNA, as shown by histone association assays that allow for the study of chromatin binding in vivo. However, Mcm10 is also required to maintain steady-state levels of DNA polymerase-α (polα). In temperature-sensitive mcm10-td mutants, depletion of Mcm10 during S phase results in degradation of the catalytic subunit of polα, without affecting other fork components such as Cdc45. We propose that Mcm10 stabilizes polα and recruits the complex to replication origins. During elongation, Mcm10 is required for the presence of polα at replication forks and may coordinate DNA synthesis with DNA unwinding by the Mcm2-7 complex.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 173-185 |
| Number of pages | 13 |
| Journal | Molecular Cell |
| Volume | 16 |
| Issue number | 2 |
| DOIs | |
| State | Published - Oct 22 2004 |
Bibliographical note
Funding Information:We thank Drs. S. Bell, J. Diffley, Y. Kawasaki, K. Labib, and B. Tye for strains and plasmids; D. Gilbert for antibodies; E. Hendrickson and D. Livingston for critical reading of the manuscript; and members of the Bielinsky lab for helpful discussions, especially S. Das-Bradoo. Special thanks go to Dr. D. Clarke for help with the fluorescence microscope. We would like to acknowledge the assistance of the Flow Cytometry Core Facility at the University of Minnesota. This work was supported by a Special Fellowship of the Leukemia and Lymphoma Society to A.-K.B. and a grant (RSG0216601) from the American Cancer Society.
