1. Dna2 is involved in CA strand resection and nascent lagging strand completion at native yeast telomeres.
- Author
-
Budd ME and Campbell JL
- Subjects
- Acetyltransferases genetics, Acetyltransferases metabolism, DNA Breaks, Double-Stranded, DNA Helicases metabolism, DNA Repair, DNA, Fungal metabolism, DNA, Single-Stranded genetics, DNA, Single-Stranded metabolism, Electrophoresis, Agar Gel, Endodeoxyribonucleases genetics, Endodeoxyribonucleases metabolism, Exodeoxyribonucleases genetics, Exodeoxyribonucleases metabolism, Flow Cytometry, Membrane Proteins genetics, Membrane Proteins metabolism, Mutation, Protein Binding, S Phase genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins metabolism, Telomere metabolism, DNA Helicases genetics, DNA, Fungal genetics, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics, Telomere genetics
- Abstract
Post-replicational telomere end processing involves both extension by telomerase and resection to produce 3'-GT-overhangs that extend beyond the complementary 5'-CA-rich strand. Resection must be carefully controlled to maintain telomere length. At short de novo telomeres generated artificially by HO endonuclease in the G2 phase, we show that dna2-defective strains are impaired in both telomere elongation and sequential 5'-CA resection. At native telomeres in dna2 mutants, GT-overhangs do clearly elongate during late S phase but are shorter than in wild type, suggesting a role for Dna2 in 5'-CA resection but also indicating significant redundancy with other nucleases. Surprisingly, elimination of Mre11 nuclease or Exo1, which are complementary to Dna2 in resection of internal double strand breaks, does not lead to further shortening of GT-overhangs in dna2 mutants. A second step in end processing involves filling in of the CA-strand to maintain appropriate telomere length. We show that Dna2 is required for normal telomeric CA-strand fill-in. Yeast dna2 mutants, like mutants in DNA ligase 1 (cdc9), accumulate low molecular weight, nascent lagging strand DNA replication intermediates at telomeres. Based on this and other results, we propose that FEN1 is not sufficient and that either Dna2 or Exo1 is required to supplement FEN1 in maturing lagging strands at telomeres. Telomeres may be among the subset of genomic locations where Dna2 helicase/nuclease is essential for the two-nuclease pathway of primer processing on lagging strands.
- Published
- 2013
- Full Text
- View/download PDF