Eukaryotic Origin-Dependent DNA Replication In Vitro Reveals Sequential Action of DDK and S-CDK Kinases
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Author(s)
Heller, Ryan C.
Kang, Sukhyun
Lam, Wendy M.
Chen, Shuyan
Chan, Clara Sophia
Bell, Stephen P
Date Issued
July 2011
Journal
Cell
Publisher
Elsevier B.V.
Citation
Heller, Ryan C., Sukhyun Kang, Wendy M. Lam, Shuyan Chen, Clara S. Chan, and Stephen P. Bell. “Eukaryotic Origin-Dependent DNA Replication In Vitro Reveals Sequential Action of DDK and S-CDK Kinases.” Cell 146, no. 1 (July 2011): 80-91. © 2011 Elsevier Inc.
Version
Final published version
Abstract
Proper eukaryotic DNA replication requires temporal separation of helicase loading from helicase activation and replisome assembly. Using an in vitro assay for eukaryotic origin-dependent replication initiation, we investigated the control of these events. After helicase loading, we found that the Dbf4-dependent Cdc7 kinase (DDK) but not S phase cyclin-dependent kinase (S-CDK) is required for the initial origin recruitment of Sld3 and the Cdc45 helicase-activating protein. Likewise, in vivo, DDK drives early-firing-origin recruitment of Cdc45 before activation of S-CDK. After S-CDK activation, a second helicase-activating protein (GINS) and the remainder of the replisome are recruited to the origin. Finally, recruitment of lagging but not leading strand DNA polymerases depends on Mcm10 and DNA unwinding. Our studies identify distinct roles for DDK and S-CDK during helicase activation and support a model in which the leading strand DNA polymerase is recruited prior to origin DNA unwinding and RNA primer synthesis.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.1016/j.cell.2011.06.012
