title: Differential requirement for SUB1 in chromosomal and plasmid double-strand DNA break repair.
abstract: Non homologous end joining (NHEJ) is an important process that repairs double strand DNA breaks (DSBs) in eukaryotic cells. Cells defective in NHEJ are unable to join chromosomal breaks. Two different NHEJ assays are typically used to determine the efficiency of NHEJ. One requires NHEJ of linearized plasmid DNA transformed into the test organism; the other requires NHEJ of a single chromosomal break induced either by HO endonuclease or the I-SceI restriction enzyme. These two assays are generally considered equivalent and rely on the same set of NHEJ genes. PC4 is an abundant DNA binding protein that has been suggested to stimulate NHEJ. Here we tested the role of PC4's yeast homolog SUB1 in repair of DNA double strand breaks using different assays. We found SUB1 is required for NHEJ repair of DSBs in plasmid DNA, but not in chromosomal DNA. Our results suggest that these two assays, while similar are not equivalent and that repair of plasmid DNA requires additional factor(s) that are not required for NHEJ repair of chromosomal double-strand DNA breaks. Possible roles for Sub1 proteins in NHEJ of plasmid DNA are discussed.
authors.affiliation: Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Lijian
authors.initials: L
authors.lastname: Yu
authors.forename: Michael R
authors.initials: MR
authors.lastname: Volkert
mesh_terms: D019457:Chromosome Breakage; D015825:Chromosomes, Fungal / Q000235:genetics / Q000378:metabolism*; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair / Q000502:physiology*; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C101206:SUB1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
references.citation: Mol Cell Biol. 2008 Aug;28(16):5082-92
references.pmid: 18541667
references.citation: EMBO J. 1996 Apr 15;15(8):1933-40
references.pmid: 8617240
references.citation: Curr Biol. 2004 Dec 14;14(23):2107-12
references.pmid: 15589152
references.citation: DNA Repair (Amst). 2010 Jun 4;9(6):617-26
references.pmid: 20356803
references.citation: Cell. 1998 Aug 7;94(3):399-409
references.pmid: 9708741
references.citation: Biochem J. 2009 Sep 25;423(2):157-68
references.pmid: 19772495
references.citation: Annu Rev Genet. 2004;38:233-71
references.pmid: 15568977
references.citation: Cell. 2004 Dec 17;119(6):777-88
references.pmid: 15607975
references.citation: Methods Enzymol. 2002;350:141-64
references.pmid: 12073310
references.citation: Cell. 2008 Sep 19;134(6):981-94
references.pmid: 18805091
references.citation: Mol Cell Biol. 2004 Jul;24(13):6084-93
references.pmid: 15199162
references.citation: Genes Dev. 2006 Sep 1;20(17):2437-49
references.pmid: 16951256
references.citation: Cell. 1994 Aug 12;78(3):513-23
references.pmid: 8062391
references.citation: Cell. 1994 Aug 12;78(3):525-34
references.pmid: 8062392
references.citation: Cell Res. 2008 Jan;18(1):114-24
references.pmid: 18166980
references.citation: EMBO J. 2005 Mar 9;24(5):1009-20
references.pmid: 15692559
references.citation: EMBO J. 1998 Sep 1;17(17):5103-11
references.pmid: 9724646
references.citation: EMBO J. 1996 Sep 16;15(18):5093-103
references.pmid: 8890183
references.citation: Annu Rev Genet. 2005;39:431-51
references.pmid: 16285867
references.citation: DNA Repair (Amst). 2007 Jul 1;6(7):923-35
references.pmid: 17363343
references.citation: Mutat Res. 2009 Oct 2;669(1-2):8-12
references.pmid: 19576233
references.citation: PLoS Genet. 2010 Feb 26;6(2):e1000855
references.pmid: 20195511
references.citation: DNA Repair (Amst). 2010 Dec 10;9(12):1256-63
references.pmid: 21115283
references.citation: DNA Repair (Amst). 2004 Jan 5;3(1):43-50
references.pmid: 14697758
references.citation: Curr Biol. 2004 Dec 14;14(23):2096-106
references.pmid: 15589151
references.citation: Cell Cycle. 2003 Sep-Oct;2(5):479-83
references.pmid: 12963848
references.citation: EMBO J. 1995 Jul 17;14(14):3520-7
references.pmid: 7628453
references.citation: Genetics. 1999 Oct;153(2):643-52
references.pmid: 10511545
references.citation: Nature. 2009 May 21;459(7245):460-3
references.pmid: 19357644
references.citation: Yeast. 1999 Jul;15(10B):963-72
references.pmid: 10407276
references.citation: Biochem Biophys Res Commun. 1999 Feb 5;255(1):88-93
references.pmid: 10082660
references.citation: Mol Cell Biol. 2005 Feb;25(3):896-906
references.pmid: 15657419
references.citation: Nature. 2001 Dec 6;414(6864):666-9
references.pmid: 11740566
references.citation: Genes Cells. 2008 Nov;13(11):1113-26
references.pmid: 18823333
references.citation: Mol Cell. 2004 Jun 4;14(5):611-23
references.pmid: 15175156
references.citation: Adv Biophys. 2004;38(Complete):21-44
references.pmid: 15476891
references.citation: Cancer Res. 2005 May 15;65(10):4020-30
references.pmid: 15899791
references.citation: Genetics. 2002 Jul;161(3):1015-27
references.pmid: 12136007
references.citation: Methods Enzymol. 2006;408:416-29
references.pmid: 16793384
references.citation: Mutat Res. 2008 Feb 1;638(1-2):56-65
references.pmid: 17928011
references.citation: Nature. 1997 Jul 31;388(6641):495-8
references.pmid: 9242411
references.citation: Biol Chem. 2002 Jun;383(6):873-92
references.pmid: 12222678
references.citation: Cell Cycle. 2011 Jan 15;10(2):261-7
references.pmid: 21212734
references.citation: Nature. 2004 Oct 21;431(7011):1011-7
references.pmid: 15496928
references.citation: Genes Dev. 1997 Aug 1;11(15):1912-24
references.pmid: 9271115
references.citation: J Mol Biol. 2009 Jan 23;385(3):788-99
references.pmid: 19038270
references.citation: Mutat Res. 2000 Jun 30;451(1-2):71-89
references.pmid: 10915866
references.citation: Nat Rev Mol Cell Biol. 2011 Feb;12(2):90-103
references.pmid: 21252998
references.citation: Nucleic Acids Res. 2000 Jul 15;28(14):2779-83
references.pmid: 10908335
references.citation: Carcinogenesis. 2002 May;23(5):687-96
references.pmid: 12016139
references.citation: Yeast. 2010 Oct;27(10):837-48
references.pmid: 20641028
references.citation: J Biol Chem. 1996 Sep 6;271(36):21842-7
references.pmid: 8702984
references.citation: Nucleic Acids Res. 1993 Jul 11;21(14):3329-30
references.pmid: 8341614
references.citation: Science. 2001 Dec 21;294(5551):2552-6
references.pmid: 11701889
references.citation: Nature. 2008 Oct 9;455(7214):770-4
references.pmid: 18806779
references.citation: Nat Rev Mol Cell Biol. 2003 Sep;4(9):712-20
references.pmid: 14506474