Article thumbnail

Sld2 binds to origin single-stranded DNA and stimulates DNA annealing

By Diane M. Kanter and Daniel L. Kaplan

Abstract

Sld2 is essential for the initiation of DNA replication, but the mechanism underlying its role in replication is not fully understood. The S-phase cyclin dependent kinase (S-CDK) triggers the association of Sld2 with Dpb11, and a phosphomimetic mutation of Sld2, Sld2T84D, functionally mimics the S-CDK phosphorylated state of Sld2. We report that Sld2T84D binds directly to the single-stranded (ss) DNA of two different origins of replication, and S-CDK phosphorylation of Sld2 stimulates the binding of Sld2 to origin ssDNA. Sld2T84D binds to a thymine-rich ssDNA region of the origin ARS1, and substitution of ARS1 thymines with adenines completely disrupts binding of Sld2T84D. Sld2T84D enhances the ability of origin ssDNA to pulldown Dpb11, and Sld2 binding to origin ssDNA may be important to allow Sld2 and Dpb11 to associate with origin DNA. We also report that Sld2T84D anneals ssDNA of an origin sequence. Dpb11 anneals ssDNA to low levels, and the addition of Sld2T84D with Dpb11 results in higher annealing activity than that of either protein alone. Sld2-stimulated annealing may be important for maintaining genome stability during the initiation of DNA replication

Topics: Genome Integrity, Repair and Replication
Publisher: Oxford University Press
OAI identifier: oai:pubmedcentral.nih.gov:3074145
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (2006). A CDK-catalysed regulatory phosphorylation for formation of the DNA replication complex Sld2–Dpb11.
  2. (1992). A yeast chromosomal origin of DNA replication defined by multiple functional elements.
  3. (2008). ATR: an essential regulator of genome integrity.
  4. (2010). CDK-dependent complex formation between replication proteins
  5. (2007). CDK-dependent phosphorylation of
  6. (1993). CLB5 and CLB6, a new pair of B cyclins involved in DNA replication in Saccharomyces cerevisiae.
  7. (2000). Dpb11 controls the association between DNA polymerases alpha and epsilon and the autonomously replicating sequence region of budding yeast.
  8. (1995). Dpb11, which interacts with DNA polymerase II(epsilon) in Saccharomyces cerevisiae, has a dual role in S-phase progression and at a cell cycle checkpoint.
  9. (1999). DRC1, DNA replication and checkpoint protein 1, functions with DPB11 to control DNA replication and the S-phase checkpoint in Saccharomyces cerevisiae.
  10. (2009). Drosophila RecQ4 has a3 0-50 DNA helicase activity that is essential for viability.
  11. (1993). Ease of DNA unwinding is a conserved property of yeast replicatiion origins.
  12. (1974). Genetic control of the cell division cycle in yeast.
  13. (2003). GINS, a novel multiprotein complex required for chromosomal DNA replication in budding yeast.
  14. (2005). Identification and functional analysis of TopBP1 and its homologs.
  15. (2007). Phosphorylation of Sld2 and Sld3 by cyclin-dependent kinases promotes DNA replication in budding yeast.
  16. (2005). Protein production by auto-induction in high-density shaking cultures.
  17. (2002). S-Cdk-dependent phosphorylation of Sld2 essential for chromosomal DNA replication in budding yeast.
  18. (1998). Sld2, which interacts with Dpb11 in Saccharomyces cerevisiae,i s required for chromosomal DNA replication.
  19. (2005). Systematic, RNA-interference-mediated identification of mus-101 modifier genes in Caenorhabditis elegans.
  20. (2006). The N-terminal noncatalytic region of Xenopus RecQ4 is required for chromatin binding of DNA polymerase alpha in the initiation of DNA replication.
  21. (2009). The S-phase checkpoint: targeting the replication fork.
  22. (1993). The structure and function of yeast ARS elements.
  23. (2002). The Xenopus Xmus101 protein is required for the recruitment of Cdc45 to origins of DNA replication.
  24. (2008). Yeast DNA replication protein Dpb11 activates the Mec1/ATR checkpoint kinase.