title: Effect of proliferating cell nuclear antigen ubiquitination and chromatin structure on the dynamic properties of the Y-family DNA polymerases.
abstract: Y-family DNA polymerases carry out translesion synthesis past damaged DNA. DNA polymerases (pol) eta and iota are usually uniformly distributed through the nucleus but accumulate in replication foci during S phase. DNA-damaging treatments result in an increase in S phase cells containing polymerase foci. Using photobleaching techniques, we show that poleta is highly mobile in human fibroblasts. Even when localized in replication foci, it is only transiently immobilized. Although ubiquitination of proliferating cell nuclear antigen (PCNA) is not required for the localization of poleta in foci, it results in an increased residence time in foci. poliota is even more mobile than poleta, both when uniformly distributed and when localized in foci. Kinetic modeling suggests that both poleta and poliota diffuse through the cell but that they are transiently immobilized for approximately 150 ms, with a larger proportion of poleta than poliota immobilized at any time. Treatment of cells with DRAQ5, which results in temporary opening of the chromatin structure, causes a dramatic immobilization of poleta but not poliota. Our data are consistent with a model in which the polymerases are transiently probing the DNA/chromatin. When DNA is exposed at replication forks, the polymerase residence times increase, and this is further facilitated by the ubiquitination of PCNA.
authors.affiliation: Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton BN1 9RQ, United Kingdom.
authors.forename: Simone
authors.initials: S
authors.lastname: Sabbioneda
authors.forename: Audrey M
authors.initials: AM
authors.lastname: Gourdin
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Green
authors.forename: Angelika
authors.initials: A
authors.lastname: Zotter
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Giglia-Mari
authors.forename: Adriaan
authors.initials: A
authors.lastname: Houtsmuller
authors.forename: Wim
authors.initials: W
authors.lastname: Vermeulen
authors.forename: Alan R
authors.initials: AR
authors.lastname: Lehmann
mesh_terms: D000818:Animals; D000880:Anthraquinones / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D006801:Humans; D009690:Nucleic Acid Conformation; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
chemical_list: D000880:Anthraquinones; D002843:Chromatin; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D018809:Proliferating Cell Nuclear Antigen; D011993:Recombinant Fusion Proteins; D000094769:DNA Polymerase iota; C520537:1,5-bis((2-(methylamino)ethyl)amino)-4,8-dihydroxyanthracene-9,10-dione; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases; C000722070:POLI protein, human
references.citation: Mol Cell. 2001 Jul;8(1):213-24
references.pmid: 11511374
references.citation: Mol Cell Biol. 2001 Nov;21(21):7199-206
references.pmid: 11585903
references.citation: Genes Dev. 2000 Jul 1;14(13):1642-50
references.pmid: 10887158
references.citation: Nature. 2002 Sep 12;419(6903):135-41
references.pmid: 12226657
references.citation: Genes Cells. 2004 Dec;9(12):1139-50
references.pmid: 15569147
references.citation: J Biol Chem. 2004 Nov 12;279(46):48360-8
references.pmid: 15342632
references.citation: Nat Cell Biol. 2006 Apr;8(4):339-47
references.pmid: 16531995
references.citation: Mol Cell Biol. 2005 Nov;25(21):9350-9
references.pmid: 16227586
references.citation: Cytometry A. 2006 Aug 1;69(8):805-14
references.pmid: 16969814
references.citation: Science. 2005 Dec 16;310(5755):1821-4
references.pmid: 16357261
references.citation: Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):7927-32
references.pmid: 10884424
references.citation: EMBO J. 2003 Mar 3;22(5):1223-33
references.pmid: 12606586
references.citation: Cell Cycle. 2004 Aug;3(8):1011-3
references.pmid: 15280666
references.citation: Histochem Cell Biol. 2001 Jan;115(1):13-21
references.pmid: 11219603
references.citation: Nature. 2004 Mar 4;428(6978):97-100
references.pmid: 14999287
references.citation: Mol Biol Cell. 2005 May;16(5):2518-28
references.pmid: 15758026
references.citation: Genes Cells. 2006 Jul;11(7):731-44
references.pmid: 16824193
references.citation: Mol Cell. 2008 Mar 14;29(5):625-36
references.pmid: 18342608
references.citation: EMBO J. 2004 Oct 1;23(19):3886-96
references.pmid: 15359278
references.citation: Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15591-8
references.pmid: 17898175
references.citation: Genes Dev. 2001 Jan 15;15(2):158-72
references.pmid: 11157773
references.citation: Methods Enzymol. 2006;408:407-15
references.pmid: 16793383
references.citation: EMBO J. 2000 Jun 15;19(12):3100-9
references.pmid: 10856253
references.citation: J Cell Biol. 2006 Apr 10;173(1):19-26
references.pmid: 16606690
references.citation: Mol Cell. 2005 May 27;18(5):499-505
references.pmid: 15916957
references.citation: EMBO J. 1998 Dec 15;17(24):7514-25
references.pmid: 9857206
references.citation: Proc Natl Acad Sci U S A. 2008 Apr 8;105(14):5361-6
references.pmid: 18385374
references.citation: Cytometry A. 2008 Jun;73(6):555-62
references.pmid: 18459157
references.citation: J Biol Chem. 1999 Jun 4;274(23):15975-7
references.pmid: 10347143
references.citation: Nature. 1976 Jun 17;261(5561):593-5
references.pmid: 934300
references.citation: Mol Cell. 2004 May 21;14(4):491-500
references.pmid: 15149598
references.citation: EMBO J. 1999 Jun 15;18(12):3491-501
references.pmid: 10369688
references.citation: Mol Cell. 2002 Dec;10(6):1355-65
references.pmid: 12504011
references.citation: Science. 1999 Jul 9;285(5425):263-5
references.pmid: 10398605