title: DNA polymerase δ stalls on telomeric lagging strand templates independently from G-quadruplex formation.
abstract: Previous evidence indicates that telomeres resemble common fragile sites and present a challenge for DNA replication. The precise impediments to replication fork progression at telomeric TTAGGG repeats are unknown, but are proposed to include G-quadruplexes (G4) on the G-rich strand. Here we examined DNA synthesis and progression by the replicative DNA polymerase δ/proliferating cell nuclear antigen/replication factor C complex on telomeric templates that mimic the leading C-rich and lagging G-rich strands. Increased polymerase stalling occurred on the G-rich template, compared with the C-rich and nontelomeric templates. Suppression of G4 formation by substituting Li(+) for K(+) as the cation, or by using templates with 7-deaza-G residues, did not alleviate Pol δ pause sites within the G residues. Furthermore, we provide evidence that G4 folding is less stable on single-stranded circular TTAGGG templates where ends are constrained, compared with linear oligonucleotides. Artificially stabilizing G4 structures on the circular templates with the G4 ligand BRACO-19 inhibited Pol δ progression into the G-rich repeats. Similar results were obtained for yeast and human Pol δ complexes. Our data indicate that G4 formation is not required for polymerase stalling on telomeric lagging strands and suggest that an alternative mechanism, in addition to stable G4s, contributes to replication stalling at telomeres.
authors.affiliation: Department of Environmental and Occupational Health, University of Pittsburgh Graduate School of Public Health, 100 Technology Drive, Pittsburgh, PA 15219, USA, Department of Physics, North Carolina State University, 2401 Stinson Drive, Raleigh, NC, 27695, USA, Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA, Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110, USA and Laboratory of Molecular Genetics and Laboratory of Structural Biology, National Institute of Environmental Health Sciences, NIH, DHHS, Research Triangle Park, NC 27709, USA.
authors.forename: Justin D
authors.initials: JD
authors.lastname: Lormand
authors.forename: Noah
authors.initials: N
authors.lastname: Buncher
authors.forename: Connor T
authors.initials: CT
authors.lastname: Murphy
authors.forename: Parminder
authors.initials: P
authors.lastname: Kaur
authors.forename: Marietta Y
authors.initials: MY
authors.lastname: Lee
authors.forename: Peter
authors.initials: P
authors.lastname: Burgers
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Kunkel
authors.forename: Patricia L
authors.initials: PL
authors.lastname: Opresko
mesh_terms: D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D004258:DNA Polymerase III / Q000378:metabolism*; D054856:G-Quadruplexes; D006801:Humans; D012091:Repetitive Sequences, Nucleic Acid; D016615:Telomere / Q000378:metabolism*; D013698:Templates, Genetic
chemical_list: D004247:DNA; D004258:DNA Polymerase III
references.citation: J Med Chem. 2009 May 14;52(9):2863-74
references.pmid: 19385599
references.citation: J Am Chem Soc. 2010 Oct 27;132(42):14910-8
references.pmid: 20882978
references.citation: Nature. 2007 Jun 21;447(7147):924-31
references.pmid: 17581575
references.citation: Science. 1998 Jan 16;279(5349):349-52
references.pmid: 9454332
references.citation: J Biol Chem. 2011 Aug 5;286(31):27132-8
references.pmid: 21676873
references.citation: Nucleic Acids Res. 2012 Aug;40(15):7358-67
references.pmid: 22576367
references.citation: Annu Rev Genet. 2007;41:169-92
references.pmid: 17608616
references.citation: Cell Biochem Biophys. 2004;41(1):75-98
references.pmid: 15371641
references.citation: J Biol Chem. 2011 Mar 4;286(9):7479-89
references.pmid: 21183684
references.citation: Nucleic Acids Res. 2001 Jul 1;29(13):2843-9
references.pmid: 11433031
references.citation: Nucleic Acids Res. 2008 Jul;36(12):4191-203
references.pmid: 18567908
references.citation: Nucleic Acids Res. 2012 Feb;40(3):1091-105
references.pmid: 21984413
references.citation: J Biol Chem. 1996 Aug 23;271(34):20958-64
references.pmid: 8702855
references.citation: Nucleic Acids Res. 2006 May 19;34(9):2723-35
references.pmid: 16714449
references.citation: J Biol Chem. 2001 May 11;276(19):16439-46
references.pmid: 11279038
references.citation: Nucleic Acids Res. 2005 Oct 12;33(18):5851-60
references.pmid: 16221978
references.citation: Nucleic Acids Res. 2008 Jan;36(1):144-56
references.pmid: 17999996
references.citation: Nucleic Acids Res. 2008 Aug;36(13):4242-56
references.pmid: 18583366
references.citation: Proc Natl Acad Sci U S A. 2005 Dec 27;102(52):18938-43
references.pmid: 16365301
references.citation: Nat Struct Mol Biol. 2009 Sep;16(9):979-86
references.pmid: 19718023
references.citation: J Biol Chem. 1994 Oct 28;269(43):27029-35
references.pmid: 7929444
references.citation: Cell. 2011 May 27;145(5):678-91
references.pmid: 21620135
references.citation: EMBO J. 2012 Jun 29;31(13):2839-51
references.pmid: 22569128
references.citation: J Biol Chem. 1993 May 15;268(14):10312-23
references.pmid: 7683674
references.citation: J Biol Chem. 1998 Jul 31;273(31):19756-62
references.pmid: 9677406
references.citation: Mol Cell. 2008 Apr 25;30(2):137-44
references.pmid: 18439893
references.citation: Mol Carcinog. 2009 Apr;48(4):286-98
references.pmid: 19123200
references.citation: Nucleic Acids Res. 2006;34(17):4857-65
references.pmid: 16973897
references.citation: Nat Struct Mol Biol. 2011 Apr;18(4):478-85
references.pmid: 21399640
references.citation: J Biol Chem. 2003 Dec 12;278(50):50537-45
references.pmid: 14523013
references.citation: Nucleic Acids Res. 2013 Feb 1;41(4):2746-55
references.pmid: 23303789
references.citation: Mol Cell. 2004 Jun 18;14(6):763-74
references.pmid: 15200954
references.citation: Nucleic Acids Res. 2006;34(19):5402-15
references.pmid: 17012276
references.citation: Science. 2004 Dec 10;306(5703):1951-3
references.pmid: 15591207
references.citation: Genes Dev. 2009 Dec 15;23(24):2915-24
references.pmid: 20008939
references.citation: Cell. 2009 Jul 10;138(1):90-103
references.pmid: 19596237
references.citation: Mech Ageing Dev. 2008 Jan-Feb;129(1-2):79-90
references.pmid: 18054793
references.citation: Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
references.pmid: 12824337
references.citation: Nucleic Acids Res. 2009 Oct;37(18):6269-75
references.pmid: 19696072
references.citation: Genes Dev. 2004 Nov 15;18(22):2764-73
references.pmid: 15520275
references.citation: Nucleic Acids Res. 2010 Mar;38(4):1149-62
references.pmid: 19969545
references.citation: Genes Dev. 2009 Sep 1;23(17):2060-75
references.pmid: 19679647
references.citation: Nat Chem. 2013 Mar;5(3):182-6
references.pmid: 23422559
references.citation: Annu Rev Genet. 2008;42:301-34
references.pmid: 18680434
references.citation: DNA Repair (Amst). 2010 Nov 10;9(11):1119-29
references.pmid: 20800555
references.citation: Cell. 2012 May 11;149(4):795-806
references.pmid: 22579284
references.citation: J Biol Chem. 2010 Aug 27;285(35):27057-27066
references.pmid: 20551483
references.citation: Nature. 2002 Jun 20;417(6891):876-80
references.pmid: 12050675
references.citation: Hum Mol Genet. 2004 Sep 1;13(17):1919-32
references.pmid: 15229185
references.citation: J Am Chem Soc. 2008 May 28;130(21):6722-4
references.pmid: 18457389
references.citation: Structure. 2012 Nov 7;20(11):1872-80
references.pmid: 22981946
references.citation: J Biol Chem. 2003 Jan 17;278(3):1618-25
references.pmid: 12424238
references.citation: Nucleic Acids Res. 2012 Feb;40(4):1636-47
references.pmid: 22021378
references.citation: Trends Biochem Sci. 2005 Jul;30(7):388-95
references.pmid: 15936947
references.citation: Biochemistry. 2008 Mar 18;47(11):3389-96
references.pmid: 18298084
references.citation: Nucleic Acids Res. 2008 Feb;36(2):688-96
references.pmid: 18079151
references.citation: Nat Rev Mol Cell Biol. 2007 Oct;8(10):825-38
references.pmid: 17885666
references.citation: Nucleic Acids Res. 2007;35(8):e58
references.pmid: 17392343
references.citation: Biochemistry. 2002 Nov 5;41(44):13133-42
references.pmid: 12403614
references.citation: DNA Repair (Amst). 2011 May 5;10(5):497-505
references.pmid: 21429821
references.citation: Nature. 2003 Nov 13;426(6963):194-8
references.pmid: 14608368
references.citation: Nature. 1990 May 31;345(6274):458-60
references.pmid: 2342578