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Proofreading activity of DNA polymerase Pol2 mediates 3'-end processing during nonhomologous end joining in yeast.
PLoS Genet. 2008 Apr 25; 4(4):e1000060.PG

Abstract

Genotoxic agents that cause double-strand breaks (DSBs) often generate damage at the break termini. Processing enzymes, including nucleases and polymerases, must remove damaged bases and/or add new bases before completion of repair. Artemis is a nuclease involved in mammalian nonhomologous end joining (NHEJ), but in Saccharomyces cerevisiae the nucleases and polymerases involved in NHEJ pathways are poorly understood. Only Pol4 has been shown to fill the gap that may form by imprecise pairing of overhanging 3' DNA ends. We previously developed a chromosomal DSB assay in yeast to study factors involved in NHEJ. Here, we use this system to examine DNA polymerases required for NHEJ in yeast. We demonstrate that Pol2 is another major DNA polymerase involved in imprecise end joining. Pol1 modulates both imprecise end joining and more complex chromosomal rearrangements, and Pol3 is primarily involved in NHEJ-mediated chromosomal rearrangements. While Pol4 is the major polymerase to fill the gap that may form by imprecise pairing of overhanging 3' DNA ends, Pol2 is important for the recession of 3' flaps that can form during imprecise pairing. Indeed, a mutation in the 3'-5' exonuclease domain of Pol2 dramatically reduces the frequency of end joins formed with initial 3' flaps. Thus, Pol2 performs a key 3' end-processing step in NHEJ.

Authors+Show Affiliations

Department of Microbiology, College of Medicine, National Taiwan University, Taipei, Taiwan.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

18437220

Citation

Tseng, Shun-Fu, et al. "Proofreading Activity of DNA Polymerase Pol2 Mediates 3'-end Processing During Nonhomologous End Joining in Yeast." PLoS Genetics, vol. 4, no. 4, 2008, pp. e1000060.
Tseng SF, Gabriel A, Teng SC. Proofreading activity of DNA polymerase Pol2 mediates 3'-end processing during nonhomologous end joining in yeast. PLoS Genet. 2008;4(4):e1000060.
Tseng, S. F., Gabriel, A., & Teng, S. C. (2008). Proofreading activity of DNA polymerase Pol2 mediates 3'-end processing during nonhomologous end joining in yeast. PLoS Genetics, 4(4), e1000060. https://doi.org/10.1371/journal.pgen.1000060
Tseng SF, Gabriel A, Teng SC. Proofreading Activity of DNA Polymerase Pol2 Mediates 3'-end Processing During Nonhomologous End Joining in Yeast. PLoS Genet. 2008 Apr 25;4(4):e1000060. PubMed PMID: 18437220.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Proofreading activity of DNA polymerase Pol2 mediates 3'-end processing during nonhomologous end joining in yeast. AU - Tseng,Shun-Fu, AU - Gabriel,Abram, AU - Teng,Shu-Chun, Y1 - 2008/04/25/ PY - 2007/08/28/received PY - 2008/03/26/accepted PY - 2008/4/26/pubmed PY - 2008/6/13/medline PY - 2008/4/26/entrez SP - e1000060 EP - e1000060 JF - PLoS genetics JO - PLoS Genet VL - 4 IS - 4 N2 - Genotoxic agents that cause double-strand breaks (DSBs) often generate damage at the break termini. Processing enzymes, including nucleases and polymerases, must remove damaged bases and/or add new bases before completion of repair. Artemis is a nuclease involved in mammalian nonhomologous end joining (NHEJ), but in Saccharomyces cerevisiae the nucleases and polymerases involved in NHEJ pathways are poorly understood. Only Pol4 has been shown to fill the gap that may form by imprecise pairing of overhanging 3' DNA ends. We previously developed a chromosomal DSB assay in yeast to study factors involved in NHEJ. Here, we use this system to examine DNA polymerases required for NHEJ in yeast. We demonstrate that Pol2 is another major DNA polymerase involved in imprecise end joining. Pol1 modulates both imprecise end joining and more complex chromosomal rearrangements, and Pol3 is primarily involved in NHEJ-mediated chromosomal rearrangements. While Pol4 is the major polymerase to fill the gap that may form by imprecise pairing of overhanging 3' DNA ends, Pol2 is important for the recession of 3' flaps that can form during imprecise pairing. Indeed, a mutation in the 3'-5' exonuclease domain of Pol2 dramatically reduces the frequency of end joins formed with initial 3' flaps. Thus, Pol2 performs a key 3' end-processing step in NHEJ. SN - 1553-7404 UR - https://www.unboundmedicine.com/medline/citation/18437220/Proofreading_activity_of_DNA_polymerase_Pol2_mediates_3'_end_processing_during_nonhomologous_end_joining_in_yeast_ L2 - https://dx.plos.org/10.1371/journal.pgen.1000060 DB - PRIME DP - Unbound Medicine ER -