title: A-type lamins maintain the positional stability of DNA damage repair foci in mammalian nuclei.
abstract: A-type lamins encoded by LMNA form a structural fibrillar meshwork within the mammalian nucleus. How this nuclear organization may influence the execution of biological processes involving DNA transactions remains unclear. Here, we characterize changes in the dynamics and biochemical interactions of lamin A/C after DNA damage. We find that DNA breakage reduces the mobility of nucleoplasmic GFP-lamin A throughout the nucleus as measured by dynamic fluorescence imaging and spectroscopy in living cells, suggestive of incorporation into stable macromolecular complexes, but does not induce the focal accumulation of GFP-lamin A at damage sites. Using a proximity ligation assay and biochemical analyses, we show that lamin A engages chromatin via histone H2AX and its phosphorylated form (γH2AX) induced by DNA damage, and that these interactions are enhanced after DNA damage. Finally, we use three-dimensional time-lapse imaging to show that LMNA inactivation significantly reduces the positional stability of DNA repair foci in living cells. This defect is partially rescued by the stable expression of GFP-lamin A. Thus collectively, our findings suggest that the dynamic structural meshwork formed by A-type lamins anchors sites of DNA repair in mammalian nuclei, providing fresh insight into the control of DNA transactions by nuclear structural organization.
authors: The Medical Research Council Cancer Cell Unit, Hutchison/MRC Research Centre, Cambridge, United Kingdom.
authors: Robert
authors: R
authors: Mahen
authors: Hiroyoshi
authors: H
authors: Hattori
authors: Miyoung
authors: M
authors: Lee
authors: Pooja
authors: P
authors: Sharma
authors: Anand D
authors: AD
authors: Jeyasekharan
authors: Ashok R
authors: AR
authors: Venkitaraman
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D006657:Histones / Q000378:metabolism; D006801:Humans; D034904:Lamin Type A / Q000378:metabolism*; D051379:Mice
chemical_list: C468783:H2AX protein, human; D006657:Histones; D034904:Lamin Type A
references: J Cell Sci. 2006 Jul 1;119(Pt 13):2704-14
references: 16772334
references: Mol Cell Biol. 2009 Feb;29(4):1050-8
references: 19064641
references: Biophys J. 2004 Jun;86(6):3473-95
references: 15189848
references: EMBO J. 2009 Aug 19;28(16):2414-27
references: 19629036
references: Proc Natl Acad Sci U S A. 2011 May 31;108(22):9310-5
references: 21576470
references: J Cell Sci. 2007 Jan 1;120(Pt 1):77-85
references: 17148572
references: Nat Cell Biol. 2012 Apr 08;14(5):510-7
references: 22484485
references: Genes Dev. 2008 Dec 15;22(24):3409-21
references: 19141474
references: Science. 2006 May 19;312(5776):1059-63
references: 16645051
references: EMBO J. 2011 Jul 12;30(16):3383-96
references: 21750527
references: J Cell Biol. 1999 Nov 29;147(5):913-20
references: 10579712
references: Nature. 2008 Nov 27;456(7221):529-33
references: 18931658
references: Nature. 2008 Nov 27;456(7221):524-8
references: 18931659
references: Cell. 2007 Feb 23;128(4):787-800
references: 17320514
references: J Cell Physiol. 1999 Mar;178(3):284-95
references: 9989774
references: Nat Methods. 2006 Dec;3(12):995-1000
references: 17072308
references: Nature. 2003 May 15;423(6937):293-8
references: 12714972
references: Nat Med. 2005 Jul;11(7):780-5
references: 15980864
references: Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2852-7
references: 10077600
references: J Cell Biol. 2000 Dec 11;151(6):1155-68
references: 11121432
references: Genes Dev. 2002 Mar 1;16(5):533-47
references: 11877373
references: J Cell Biol. 2007 Jul 16;178(2):209-18
references: 17635934
references: Cold Spring Harb Symp Quant Biol. 2010;75:533-43
references: 21209392
references: Nature. 2008 Jun 12;453(7197):948-51
references: 18463634
references: Nat Cell Biol. 2007 Jun;9(6):675-82
references: 17486118
references: J Cell Sci. 1995 Feb;108 ( Pt 2):635-44
references: 7769007
references: Cell. 2011 Mar 4;144(5):732-44
references: 21353298
references: J Cell Sci. 1999 Oct;112 ( Pt 20):3463-75
references: 10504295
references: Nature. 2007 May 24;447(7143):413-7
references: 17522674
references: IEEE Trans Image Process. 1998;7(1):27-41
references: 18267377
references: Science. 2004 Jan 2;303(5654):92-5
references: 14704429
references: Methods Enzymol. 2004;375:393-414
references: 14870680
references: Nature. 1986 Oct 9-15;323(6088):560-4
references: 3762708
references: Nucleus. 2012 Sep-Oct;3(5):463-74
references: 22895093
references: Proc Natl Acad Sci U S A. 2006 Jul 5;103(27):10271-10276
references: 16801550
references: Nat Cell Biol. 2012 Apr 08;14(5):502-9
references: 22484486
references: J Biol Chem. 1998 Mar 6;273(10):5858-68
references: 9488723
references: Exp Cell Res. 2009 Apr 1;315(6):996-1007
references: 19272320