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Pol3 is involved in nonhomologous end-joining in Saccharomyces cerevisiae.
DNA Repair (Amst). 2008 Sep 01; 7(9):1531-41.DR

Abstract

Nonhomologous end joining connects DNA ends in the absence of extended sequence homology and requires removal of mismatched DNA ends and gap-filling synthesis prior to a religation step. Pol4 within the Pol X family is the only polymerase known to be involved in end processing during nonhomologous end joining in yeast. The Saccharomyces cerevisiae POL3/CDC2 gene encodes polymerase delta that is involved in DNA replication and other DNA repair processes. Here, we show that POL3 is involved in nonhomologous end joining using a plasmid-based end-joining assay in yeast, in which the pol3-t mutation caused a 1.9- to 3.2-fold decrease in the end-joining efficiency of partially compatible 5' or 3' ends, or incompatible ends, similar to the pol4 mutant. The pol3-t pol4 double mutation showed a synergistic decrease in the efficiency of NHEJ with partially compatible 5' ends or incompatible ends. Sequence analysis of the rejoined junctions recovered from the wild-type cells and mutants indicated that POL3 is required for gap filling at 3' overhangs, but not 5' overhangs during POL4-independent nonhomologous end joining. We also show that either Pol3 or Pol4 is required for simple religation of compatible or blunt ends. These results suggest that Pol3 has a generalized function in end joining in addition to its role in gap filling at 3' overhangs to enhance the overall efficiency of nonhomologous end joining. Moreover, the decreased end-joining efficiency seen in the pol3-t mutant was not due to S-phase arrest associated with the mutant. Taken together, our genetic evidence supports a novel role of Pol3 in nonhomologous end joining that facilitates gap filling at 3' overhangs in the absence of Pol4 to maintain genomic integrity.

Authors+Show Affiliations

Department of Pathology and Radiation Oncology, David Geffen School of Medicine at UCLA, University of California-Los Angeles, 650 Charles E. Young Drive South, Los Angeles, CA 90095, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18606574

Citation

Chan, Cecilia Y., et al. "Pol3 Is Involved in Nonhomologous End-joining in Saccharomyces Cerevisiae." DNA Repair, vol. 7, no. 9, 2008, pp. 1531-41.
Chan CY, Galli A, Schiestl RH. Pol3 is involved in nonhomologous end-joining in Saccharomyces cerevisiae. DNA Repair (Amst). 2008;7(9):1531-41.
Chan, C. Y., Galli, A., & Schiestl, R. H. (2008). Pol3 is involved in nonhomologous end-joining in Saccharomyces cerevisiae. DNA Repair, 7(9), 1531-41. https://doi.org/10.1016/j.dnarep.2008.05.008
Chan CY, Galli A, Schiestl RH. Pol3 Is Involved in Nonhomologous End-joining in Saccharomyces Cerevisiae. DNA Repair (Amst). 2008 Sep 1;7(9):1531-41. PubMed PMID: 18606574.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Pol3 is involved in nonhomologous end-joining in Saccharomyces cerevisiae. AU - Chan,Cecilia Y, AU - Galli,Alvaro, AU - Schiestl,Robert H, Y1 - 2008/07/07/ PY - 2007/12/16/received PY - 2008/05/14/revised PY - 2008/05/19/accepted PY - 2008/7/9/pubmed PY - 2008/12/24/medline PY - 2008/7/9/entrez SP - 1531 EP - 41 JF - DNA repair JO - DNA Repair (Amst) VL - 7 IS - 9 N2 - Nonhomologous end joining connects DNA ends in the absence of extended sequence homology and requires removal of mismatched DNA ends and gap-filling synthesis prior to a religation step. Pol4 within the Pol X family is the only polymerase known to be involved in end processing during nonhomologous end joining in yeast. The Saccharomyces cerevisiae POL3/CDC2 gene encodes polymerase delta that is involved in DNA replication and other DNA repair processes. Here, we show that POL3 is involved in nonhomologous end joining using a plasmid-based end-joining assay in yeast, in which the pol3-t mutation caused a 1.9- to 3.2-fold decrease in the end-joining efficiency of partially compatible 5' or 3' ends, or incompatible ends, similar to the pol4 mutant. The pol3-t pol4 double mutation showed a synergistic decrease in the efficiency of NHEJ with partially compatible 5' ends or incompatible ends. Sequence analysis of the rejoined junctions recovered from the wild-type cells and mutants indicated that POL3 is required for gap filling at 3' overhangs, but not 5' overhangs during POL4-independent nonhomologous end joining. We also show that either Pol3 or Pol4 is required for simple religation of compatible or blunt ends. These results suggest that Pol3 has a generalized function in end joining in addition to its role in gap filling at 3' overhangs to enhance the overall efficiency of nonhomologous end joining. Moreover, the decreased end-joining efficiency seen in the pol3-t mutant was not due to S-phase arrest associated with the mutant. Taken together, our genetic evidence supports a novel role of Pol3 in nonhomologous end joining that facilitates gap filling at 3' overhangs in the absence of Pol4 to maintain genomic integrity. SN - 1568-7864 UR - https://www.unboundmedicine.com/medline/citation/18606574/Pol3_is_involved_in_nonhomologous_end_joining_in_Saccharomyces_cerevisiae_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1568-7864(08)00194-8 DB - PRIME DP - Unbound Medicine ER -