title: 5'CAG and 5'CTG Repeats Create Differential Impediment to the Progression of a Minimal Reconstituted T4 Replisome Depending on the Concentration of dNTPs.
abstract: Instability of repetitive sequences originates from strand misalignment during repair or replicative DNA synthesis. To investigate the activity of reconstituted T4 replisomes across trinucleotide repeats (TNRs) during leading strand DNA synthesis, we developed a method to build replication miniforks containing a TNR unit of defined sequence and length. Each minifork consists of three strands, primer, leading strand template, and lagging strand template with a 5' single-stranded (ss) tail. Each strand is prepared independently, and the minifork is assembled by hybridization of the three strands. Using these miniforks and a minimal reconstituted T4 replisome, we show that during leading strand DNA synthesis, the dNTP concentration dictates which strand of the structure-forming 5'CAG/5'CTG repeat creates the strongest impediment to the minimal replication complex. We discuss this result in the light of the known fluctuation of dNTP concentration during the cell cycle and cell growth and the known concentration balance among individual dNTPs.
authors.affiliation: Muséum National d'Histoire Naturelle, Département "Régulations, Développement et Diversité Moléculaire", Laboratoire de Régulations et Dynamique des Génomes, USM 0503-INSERM U 565-UMR 7196, Case Postale no 26, 57 rue Cuvier, 75231 Paris cedex 05, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Delagoutte
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Baldacci
references.citation: Nucleic Acids Res. 2010 Apr;38(6):e85
references.pmid: 20008099
references.citation: Biochemistry. 1992 Sep 22;31(37):8675-90
references.pmid: 1390652
references.citation: Virol J. 2010 Dec 03;7:359
references.pmid: 21129204
references.citation: Proc Natl Acad Sci U S A. 2007 Jan 23;104(4):1183-8
references.pmid: 17227840
references.citation: J Biol Chem. 2008 May 9;283(19):13341-56
references.pmid: 18263578
references.citation: Virol J. 2010 Dec 03;7:358
references.pmid: 21129203
references.citation: Adv Exp Med Biol. 2010;685:45-63
references.pmid: 20687494
references.citation: J Biol Chem. 1979 Jul 25;254(14):6797-802
references.pmid: 376535
references.citation: Trends Genet. 2000 Dec;16(12):551-8
references.pmid: 11102705
references.citation: Nat Chem Biol. 2010 Sep;6(9):652-9
references.pmid: 20676085
references.citation: Biochim Biophys Acta. 1972 May 10;269(2):184-91
references.pmid: 4555254
references.citation: FASEB J. 1992 Feb 1;6(3):871-8
references.pmid: 1310946
references.citation: Science. 2009 May 29;324(5931):1213-6
references.pmid: 19478187
references.citation: Ann Neurol. 2010 Mar;67(3):291-300
references.pmid: 20373340
references.citation: Antioxid Redox Signal. 2010 Sep 1;13(5):651-90
references.pmid: 20156111
references.citation: J Biol Chem. 1979 Jul 25;254(14):6787-96
references.pmid: 376534
references.citation: Cancer Res. 2002 May 1;62(9):2447-54
references.pmid: 11980631
references.citation: Biochim Biophys Acta. 2010 May;1804(5):1094-112
references.pmid: 19665592
references.citation: Nature. 2002 Feb 21;415(6874):910-3
references.pmid: 11859369
references.citation: Annu Rev Biochem. 2001;70:181-208
references.pmid: 11395406
references.citation: J Bacteriol. 1989 May;171(5):2265-70
references.pmid: 2651395
references.citation: J Biol Chem. 1989 Mar 15;264(8):4725-31
references.pmid: 2538456
references.citation: Science. 1994 Feb 11;263(5148):808-11
references.pmid: 8303297
references.citation: Cell. 2003 Feb 7;112(3):391-401
references.pmid: 12581528
references.citation: Science. 2005 Jun 10;308(5728):1630-4
references.pmid: 15947188
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 28;101(52):18058-63
references.pmid: 15596718
references.citation: Med Sci (Paris). 2003 Jan;19(1):55-62
references.pmid: 12836192
references.citation: J Bacteriol. 2008 Jan;190(2):718-26
references.pmid: 17965154
references.citation: Nucleic Acids Res. 2010 Jul;38(12):3975-83
references.pmid: 20215435
references.citation: Methods Enzymol. 1995;262:560-84
references.pmid: 8594379
references.citation: Trends Neurosci. 2010 Jun;33(6):292-8
references.pmid: 20398949
references.citation: J Biol Chem. 1994 Dec 30;269(52):33069-81
references.pmid: 7806535
references.citation: Nat Rev Genet. 2010 Nov;11(11):786-99
references.pmid: 20953213