Phosphatase complex Pph3/Psy2 is involved in regulation of efficient non-homologous end-joining pathway in the yeast Saccharomyces cerevisiae

Katayoun Omidi; Mohsen Hooshyar; Matthew Jessulat; Bahram Samanfar; Megan Sanders; Daniel Burnside; Sylvain Pitre; Andrew Schoenrock; Jianhua Xu; Mohan Babu; Ashkan Golshani

doi:10.1371/journal.pone.0087248

Phosphatase complex Pph3/Psy2 is involved in regulation of efficient non-homologous end-joining pathway in the yeast Saccharomyces cerevisiae

PLoS One. 2014 Jan 31;9(1):e87248. doi: 10.1371/journal.pone.0087248. eCollection 2014.

Authors

Affiliations

¹ Department of Biology, Carleton University, Ottawa, Ontario, Canada ; Ottawa Institute of Systems Biology, Carleton University, Ottawa, Ontario, Canada.
² Department of Biology, Carleton University, Ottawa, Ontario, Canada ; Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
³ Department of Computer Science, Carleton University, Ottawa, Ontario, Canada.
⁴ College of Pharmaceutical Sciences, Zhejian University, Hangzhou, Zhejiang, China.
⁵ Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.

Abstract

One of the main mechanisms for double stranded DNA break (DSB) repair is through the non-homologous end-joining (NHEJ) pathway. Using plasmid and chromosomal repair assays, we showed that deletion mutant strains for interacting proteins Pph3p and Psy2p had reduced efficiencies in NHEJ. We further observed that this activity of Pph3p and Psy2p appeared linked to cell cycle Rad53p and Chk1p checkpoint proteins. Pph3/Psy2 is a phosphatase complex, which regulates recovery from the Rad53p DNA damage checkpoint. Overexpression of Chk1p checkpoint protein in a parallel pathway to Rad53p compensated for the deletion of PPH3 or PSY2 in a chromosomal repair assay. Double mutant strains Δpph3/Δchk1 and Δpsy2/Δchk1 showed additional reductions in the efficiency of plasmid repair, compared to both single deletions which is in agreement with the activity of Pph3p and Psy2p in a parallel pathway to Chk1p. Genetic interaction analyses also supported a role for Pph3p and Psy2p in DNA damage repair, the NHEJ pathway, as well as cell cycle progression. Collectively, we report that the activity of Pph3p and Psy2p further connects NHEJ repair to cell cycle progression.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Cycle / genetics
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism
Checkpoint Kinase 1
Checkpoint Kinase 2 / genetics
Checkpoint Kinase 2 / metabolism
DNA Breaks, Double-Stranded
DNA End-Joining Repair*
Multiprotein Complexes / genetics
Multiprotein Complexes / metabolism
Mutation
Nuclear Proteins / genetics
Nuclear Proteins / metabolism*
Phosphoprotein Phosphatases / genetics
Phosphoprotein Phosphatases / metabolism*
Protein Binding
Protein Interaction Mapping
Protein Kinases / genetics
Protein Kinases / metabolism
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism*

Substances

Cell Cycle Proteins
Multiprotein Complexes
Nuclear Proteins
Psy2 protein, S cerevisiae
Saccharomyces cerevisiae Proteins
Protein Kinases
Checkpoint Kinase 2
Checkpoint Kinase 1
RAD53 protein, S cerevisiae
PPH3 protein, S cerevisiae
Phosphoprotein Phosphatases

Grants and funding

This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.