Genetic and physical interactions between DPB11 and DDC1 in the yeast DNA damage response pathway

Hong Wang, Stephen J. Elledge

Research output: Contribution to journalArticlepeer-review

68 Scopus citations

Abstract

DPB11 is essential for DNA replication and S/M checkpoint control in Saccharomyces cerevisiae. The Dpb11 protein contains four BRCT domains, which have been proposed to be involved in protein-protein interactions. To further investigate the regulation and function of Dpb11, a yeast two-hybrid screen was carried out to identify proteins that physically interact with Dpb11. One positive clone isolated from the screen encoded a carboxyl-terminal fragment of Ddc1 (339-612 aa). Ddc1 is a DNA damage checkpoint protein, which, together with Mec3 and Rad17, has been proposed to form a PCNA-like complex and acts upstream in the DNA damage checkpoint pathways. We further determined that the carboxyl region of Dpb11 is required for its interaction with Ddc1. DDC1 and DPB11 also interact genetically. The Δddc1 dpb11-1 double mutant is more UV and MMS sensitive than the Δddc1 or the dpb11-1 single mutants. Furthermore, the double mutant is more hydroxyurea sensitive and displayed a lower restrictive temperature than dpb11-1. These results suggest that DPB11 and DDC1 may function in the same or parallel pathways after DNA damage and that DDC1 may play a role in responding to replication defects.

Original languageEnglish
Pages (from-to)1295-1304
Number of pages10
JournalGenetics
Volume160
Issue number4
DOIs
StatePublished - 2002

Keywords

  • Cell Cycle Proteins/genetics
  • DNA Repair/genetics
  • DNA, Fungal/physiology
  • In Vitro Techniques
  • Phosphoproteins/genetics
  • Saccharomyces cerevisiae Proteins/genetics
  • Saccharomyces cerevisiae/genetics
  • Two-Hybrid System Techniques

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