Deletion of yeast TPK1 reduces the efficiency of non-homologous end joining DNA repair

Mohsen Hooshyar; Matthew Jessulat; Daniel Burnside; Anna Kluew; Mohan Babu; Ashkan Golshani

doi:10.1016/j.bbrc.2020.09.083

Deletion of yeast TPK1 reduces the efficiency of non-homologous end joining DNA repair

Biochem Biophys Res Commun. 2020 Dec 17;533(4):899-904. doi: 10.1016/j.bbrc.2020.09.083. Epub 2020 Sep 30.

Authors

Mohsen Hooshyar¹, Matthew Jessulat², Daniel Burnside¹, Anna Kluew³, Mohan Babu⁴, Ashkan Golshani⁵

Affiliations

¹ Department of Biology, Carleton University, Ottawa, Ontario, Canada; Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Ontario, Canada.
² Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada.
³ Department of Biology, Carleton University, Ottawa, Ontario, Canada.
⁴ Department of Biochemistry, Research and Innovation Centre, University of Regina, Regina, Saskatchewan, Canada. Electronic address: mohan.babu@uregina.ca.
⁵ Department of Biology, Carleton University, Ottawa, Ontario, Canada; Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Ontario, Canada. Electronic address: Ashkan.Golshani@carleton.ca.

PMID: 33008596
DOI: 10.1016/j.bbrc.2020.09.083

Abstract

Non-homologous end joining (NHEJ) is a highly conserved mechanism of DNA double-stranded break (DSB) repair. Here we utilize a computational protein-protein interaction method to identify human PRKACB as a potential candidate interacting with NHEJ proteins. We show that the deletion of its yeast homolog, TPK1 that codes for the protein kinase A catalytic subunit reduces the efficiency of NHEJ repair of breaks with overhangs and blunt ends in plasmid-based repair assays. Additionally, tpk1Δ mutants showed defects in the repair of chromosomal breaks induced by HO-site specific endonuclease. Our double deletion mutant analyses suggest that TPK1 and YKU80, a key player in NHEJ could function in parallel pathways. Altogether, here we report a novel involvement for TPK1 in NHEJ.

Keywords: DNA double Stranded breaks; DNA repair; Non-homologous end joining; TPK1; YKU80; Yeast.

Publication types

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

MeSH terms

Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / genetics
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / metabolism
Cyclic AMP-Dependent Protein Kinases / deficiency
Cyclic AMP-Dependent Protein Kinases / genetics*
Cyclic AMP-Dependent Protein Kinases / metabolism*
DNA Breaks, Double-Stranded
DNA End-Joining Repair / genetics*
DNA End-Joining Repair / physiology*
DNA, Fungal / genetics
DNA, Fungal / metabolism
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Gene Deletion
Genes, Fungal
Genes, Synthetic
Genetic Association Studies
Humans
Protein Interaction Maps
Saccharomyces cerevisiae / genetics*
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / genetics*
Saccharomyces cerevisiae Proteins / metabolism*

Substances

DNA, Fungal
DNA-Binding Proteins
Saccharomyces cerevisiae Proteins
YKU80 protein, S cerevisiae
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
Cyclic AMP-Dependent Protein Kinases
PRKACB protein, human
Tpk1 protein, S cerevisiae