Disruption of PCNA-lamins A/C interactions by prelamin A induces DNA replication fork stalling.
The accumulation of prelamin A is linked to disruption of cellular homeostasis, tissue degeneration and aging. Its expression is implicated in compromised genome stability and increased levels of DNA damage, but to date there is no complete explanation for how prelamin A exerts its toxic effects. As the nuclear lamina is important for DNA replication we wanted to investigate the relationship between prelamin A expression and DNA replication fork stability. In this study we report that the expression of prelamin A in U2OS cells induced both mono-ubiquitination of proliferating cell nuclear antigen (PCNA) and subsequent induction of Pol η, two hallmarks of DNA replication fork stalling. Immunofluorescence microscopy revealed that cells expressing prelamin A presented with high levels of colocalisation between PCNA and γH2AX, indicating collapse of stalled DNA replication forks into DNA double-strand breaks. Subsequent protein-protein interaction assays showed prelamin A interacted with PCNA and that its presence mitigated interactions between PCNA and the mature nuclear lamina. Thus, we propose that the cytotoxicity of prelamin A arises in part, from it actively competing against mature lamin A to bind PCNA and that this destabilises DNA replication to induce fork stalling which in turn contributes to genomic instability.
a King's College London , The James Black Center , London , United Kingdom.
Andrew M
AM
Cobb
a King's College London , The James Black Center , London , United Kingdom.
Thomas V
TV
Murray
a King's College London , The James Black Center , London , United Kingdom.
Derek T
DT
Warren
a King's College London , The James Black Center , London , United Kingdom.
Yiwen
Y
Liu
a King's College London , The James Black Center , London , United Kingdom.
Catherine M
CM
Shanahan
D045744:Cell Line, Tumor; D004249:DNA Damage; D004261:DNA Replication; D019281:Dimerization; D006657:Histones / Q000378:metabolism; D006801:Humans; D034904:Lamin Type A / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D013941:Thymine / Q000378:metabolism; D054875:Ubiquitination
C468783:H2AX protein, human; D006657:Histones; D034904:Lamin Type A; D018809:Proliferating Cell Nuclear Antigen; C090764:prelamin A; D013941:Thymine
DNA damage; DNA replication fork stalling; PCNA; Prelamin A; nuclear lamina
J Cell Biol. 1994 Jun;125(6):1201-12
7911470
Oncogene. 2004 Jan 8;23(1):201-12
14712225
Hum Mol Genet. 2005 Jun 1;14(11):1503-13
15843403
Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9677-82
15210943
Exp Cell Res. 2010 Oct 15;316(17):2747-59
20599958
Mol Cell. 2004 May 21;14 (4):491-500
15149598
Exp Cell Res. 2008 Feb 1;314(3):453-62
18093584
Nature. 2003 May 15;423(6937):293-8
12714972
FEBS Lett. 2011 Sep 16;585(18):2780-5
21640107
Proc Natl Acad Sci U S A. 2014 Jun 3;111(22):E2261-70
24843141
J Invest Dermatol. 2013 Jul;133(7):1857-62
23392295
Biochem Cell Biol. 2011 Feb;89(1):45-60
21326362
Microbiol Mol Biol Rev. 2007 Mar;71(1):13-35
17347517
Genes Cells. 2008 Apr;13(4):343-54
18363965
Int J Mol Sci. 2012;13(9):11844-60
23109886
J Biol Chem. 2012 Sep 28;287(40):33327-38
22887995
Nat Med. 2005 Jul;11(7):780-5
15980864
Mol Cell Biol. 2013 Mar;33(6):1210-22
23319047
PLoS One. 2007 Oct 24;2(10):e1057
17957241
Proc Natl Acad Sci U S A. 1990 Aug;87(15):5672-6
1974050
Genes Dev. 2000 Nov 15;14(22):2855-68
11090133
Biochim Biophys Acta. 2008 Dec;1783(12):2398-414
18706456
EMBO J. 2004 Oct 1;23 (19):3886-96
15359278
Photochem Photobiol Sci. 2012 Jan;11(1):74-80
21901217
Proc Natl Acad Sci U S A. 2006 Feb 28;103(9):3250-5
16492728
Novartis Found Symp. 2007;283:174-91; discussion 191-3, 238-41
18300422
FASEB J. 2008 Feb;22(2):603-11
17848622
EMBO Rep. 2007 Apr;8(4):346-53
17401409
Biochem Soc Trans. 2011 Dec;39(6):1764-9
22103522
Circulation. 2010 May 25;121(20):2200-10
20458013
Nucleic Acids Res. 1995 Oct 25;23(20):4202-9
7479085
Genes Dev. 2008 Apr 1;22(7):832-53
18381888
J Cell Biol. 1997 Mar 24;136(6):1201-12
9087437
Genome Res. 2013 Feb;23(2):260-9
23152449
J Cell Sci. 2006 Jul 1;119(Pt 13):2704-14
16772334
Nat Rev Mol Cell Biol. 2005 Jan;6(1):21-31
15688064
Biochem Soc Trans. 2011 Dec;39(6):1780-5
22103525
J Cell Biol. 2008 Apr 21;181(2):269-80
18426975
EMBO J. 2006 Jun 21;25(12):2847-55
16763556
Aging Cell. 2016 Jul 27;:
27464478
Ageing Res Rev. 2017 Jan;33:18-29
27374873
J Cell Biochem. 2007 Dec 1;102(5):1149-59
17654502
Arterioscler Thromb Vasc Biol. 2010 Nov;30(11):2301-9
20798379
Mol Biol Cell. 2014 May;25(9):1493-510
24623722
Glob Heart. 2014 Jun;9(2):211-8
25667091
Science. 2006 May 19;312(5776):1059-63
16645051
J Photochem Photobiol B. 2001 Oct;63(1-3):88-102
11684456
J Cell Sci. 2006 Aug 15;119(Pt 16):3265-72
16899817
Nucleus. 2011 Jan-Feb;2(1):4-9
21647293
Aging (Albany NY). 2009 Jan;1(1):28-37
19851476
J Biol Chem. 2010 Jan 8;285(2):1311-20
19861416
J Cell Biol. 2000 Jun 12;149(6):1179-92
10851016
Mol Cell Biol. 2004 Jan;24(2):595-607
14701733
Nucleus. 2015;6(3):236-46
25996284
PLoS One. 2013 May 02;8(5):e61893
23658700