Lamin A/C Depletion Enhances DNA Damage-Induced Stalled Replication Fork Arrest

ABSTRACT The human LMNA gene encodes the essential nuclear envelope proteins lamin A and C (lamin A/C). Mutations in LMNA result in altered nuclear morphology, but how this impacts the mechanisms that maintain genomic stability is unclear. Here, we report that lamin A/C-deficient cells have a normal response to ionizing radiation but are sensitive to agents that cause interstrand cross-links (ICLs) or replication stress. In response to treatment with ICL agents (cisplatin, camptothecin, and mitomycin), lamin A/C-deficient cells displayed normal γ-H2AX focus formation but a higher frequency of cells with delayed γ-H2AX removal, decreased recruitment of the FANCD2 repair factor, and a higher frequency of chromosome aberrations. Similarly, following hydroxyurea-induced replication stress, lamin A/C-deficient cells had an increased frequency of cells with delayed disappearance of γ-H2AX foci and defective repair factor recruitment (Mre11, CtIP, Rad51, RPA, and FANCD2). Replicative stress also resulted in a higher frequency of chromosomal aberrations as well as defective replication restart. Taken together, the data can be interpreted to suggest that lamin A/C has a role in the restart of stalled replication forks, a prerequisite for initiation of DNA damage repair by the homologous recombination pathway, which is intact in lamin A/C-deficient cells. We propose that lamin A/C is required for maintaining genomic stability following replication fork stalling, induced by either ICL damage or replicative stress, in order to facilitate fork regression prior to DNA damage repair.

[1]  Benjamin P. C. Chen,et al.  Lamin A/C Depletion Enhances DNA Damage-Induced Stalled Replication Fork Arrest , 2013, Molecular and Cellular Biology.

[2]  Y. Zou,et al.  DNA-damage accumulation and replicative arrest in Hutchinson-Gilford progeria syndrome. , 2011, Biochemical Society transactions.

[3]  Wei Yang,et al.  Structure and mechanism of human DNA polymerase η , 2011, Nature.

[4]  Joshua E. Elias,et al.  A function for cyclin D1 in DNA repair uncovered by protein interactome analyses in human cancers , 2011, Nature.

[5]  A. Egashira,et al.  Double-Strand Break Repair-Independent Role for BRCA2 in Blocking Stalled Replication Fork Degradation by MRE11 , 2011, Cell.

[6]  E. McNally,et al.  Gene expression, chromosome position and lamin A/C mutations , 2011 .

[7]  T. Helleday,et al.  Pathways of mammalian replication fork restart , 2010, Nature Reviews Molecular Cell Biology.

[8]  Robert E. Johnson,et al.  Structural basis for the suppression of skin cancers by DNA polymerase η , 2010, Nature.

[9]  Jae Young Lee,et al.  Structure and Mechanism of Human DNA Polymerase η , 2010, Nature.

[10]  David J. Chen,et al.  MOF and Histone H4 Acetylation at Lysine 16 Are Critical for DNA Damage Response and Double-Strand Break Repair , 2010, Molecular and Cellular Biology.

[11]  T. Helleday,et al.  Hydroxyurea-Stalled Replication Forks Become Progressively Inactivated and Require Two Different RAD51-Mediated Pathways for Restart and Repair , 2010, Molecular cell.

[12]  T. Pandita,et al.  Cell cycle checkpoint defects contribute to genomic instability in PTEN deficient cells independent of DNA DSB repair , 2009, Cell cycle.

[13]  Jingchuan Sun,et al.  Mechanism of Replication-Coupled DNA Interstrand Crosslink Repair , 2008, Cell.

[14]  S. Adam,et al.  Nuclear lamins and chromatin: when structure meets function. , 2009, Advances in enzyme regulation.

[15]  Thomas Cremer,et al.  The A- and B-type nuclear lamin networks: microdomains involved in chromatin organization and transcription. , 2008, Genes & development.

[16]  E. C. Schirmer.  The epigenetics of nuclear envelope organization and disease. , 2008, Mutation research.

[17]  T. Pandita,et al.  Inhibition of telomerase activity enhances hyperthermia-mediated radiosensitization. , 2008, Cancer research.

[18]  M. C. Cardoso,et al.  The highly conserved nuclear lamin Ig-fold binds to PCNA: its role in DNA replication , 2008, The Journal of cell biology.

[19]  Kaushik Sengupta,et al.  Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin. , 2008, Genes & development.

[20]  T. Ludwig,et al.  The Mammalian Ortholog of Drosophila MOF That Acetylates Histone H4 Lysine 16 Is Essential for Embryogenesis and Oncogenesis , 2007, Molecular and Cellular Biology.

[21]  S. Young,et al.  Mouse models of the laminopathies. , 2007, Experimental cell research.

[22]  M. Löbrich,et al.  Hyperthermia activates a subset of ataxia-telangiectasia mutated effectors independent of DNA strand breaks and heat shock protein 70 status. , 2007, Cancer research.

[23]  Richard T. Lee,et al.  Lamins A and C but Not Lamin B1 Regulate Nuclear Mechanics* , 2006, Journal of Biological Chemistry.

[24]  J. Broers,et al.  Nuclear lamins: laminopathies and their role in premature ageing. , 2006, Physiological reviews.

[25]  T. Pandita.  Role of Mammalian Rad9 in Genomic Stability and Ionizing Radiation Response , 2006, Cell cycle.

[26]  T. Pandita,et al.  Mammalian Rad9 Plays a Role in Telomere Stability, S- and G2-Phase-Specific Cell Survival, and Homologous Recombinational Repair , 2006, Molecular and Cellular Biology.

[27]  S. Lees-Miller.  Dysfunction of lamin A triggers a DNA damage response and cellular senescence. , 2006, DNA repair.

[28]  T. Ludwig,et al.  Involvement of Human MOF in ATM Function , 2005, Molecular and Cellular Biology.

[29]  Katherine L. Wilson,et al.  The nuclear lamina comes of age , 2005, Nature Reviews Molecular Cell Biology.

[30]  Florence Hediger,et al.  The function of nuclear architecture: a genetic approach. , 2004, Annual review of genetics.

[31]  R. Hennekam,et al.  Lamin A and ZMPSTE24 (FACE-1) defects cause nuclear disorganization and identify restrictive dermopathy as a lethal neonatal laminopathy. , 2004, Human molecular genetics.

[32]  T. Pandita,et al.  Human Heterochromatin Protein 1 Isoforms HP1Hsα and HP1Hsβ Interfere with hTERT-Telomere Interactions and Correlate with Changes in Cell Growth and Response to Ionizing Radiation , 2003, Molecular and Cellular Biology.

[33]  Penny A. Johnson,et al.  XRCC3 and Rad51 modulate replication fork progression on damaged vertebrate chromosomes. , 2003, Molecular cell.

[34]  H. Scherthan,et al.  hTERT associates with human telomeres and enhances genomic stability and DNA repair , 2003, Oncogene.

[35]  C. Stewart,et al.  Life at the edge: the nuclear envelope and human disease , 2002, Nature Reviews Molecular Cell Biology.

[36]  G. Wahl,et al.  c-Myc can induce DNA damage, increase reactive oxygen species, and mitigate p53 function: a mechanism for oncogene-induced genetic instability. , 2002, Molecular cell.

[37]  R. Goldman,et al.  Nuclear Lamins a and B1: Different Pathways of Assembly during Nuclear Envelope Formation in Living Cells , 2000 .

[38]  B. Kennedy,et al.  Nuclear organization of DNA replication in primary mammalian cells. , 2000, Genes & development.

[39]  T. Pandita,et al.  Ionizing radiation activates the ATM kinase throughout the cell cycle , 2000, Oncogene.

[40]  Brian Burke,et al.  Loss of a-Type Lamin Expression Compromises Nuclear Envelope Integrity Leading to Muscular Dystrophy , 1999, The Journal of cell biology.

[41]  T. Pandita,et al.  Altered Telomere Nuclear Matrix Interactions and Nucleosomal Periodicity in Ataxia Telangiectasia Cells before and after Ionizing Radiation Treatment , 1999, Molecular and Cellular Biology.

[42]  Harry Scherthan,et al.  Atm Inactivation Results in Aberrant Telomere Clustering during Meiotic Prophase , 1999, Molecular and Cellular Biology.

[43]  T. Richmond,et al.  The tail domain of lamin Dm0 binds histones H2A and H2B. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Ana Pombo,et al.  Replicon Clusters Are Stable Units of Chromosome Structure: Evidence That Nuclear Organization Contributes to the Efficient Activation and Propagation of S Phase in Human Cells , 1998, The Journal of cell biology.

[45]  S. Lavi,et al.  Induction of circles of heterogeneous sizes in carcinogen-treated cells: two-dimensional gel analysis of circular DNA molecules , 1996, Molecular and cellular biology.

[46]  T. Pandita,et al.  Chromosome end associations, telomeres and telomerase activity in ataxia telangiectasia cells. , 1995, Cytogenetics and cell genetics.

[47]  T. Pandita,et al.  Spontaneous amplification of interstitial telomeric bands in Chinese hamster ovary cells. , 1995, Cytogenetics and cell genetics.

[48]  T. de Lange.  Human telomeres are attached to the nuclear matrix. , 1992, The EMBO journal.

[49]  T. Pandita,et al.  The contribution of DNA and chromosome repair deficiencies to the radiosensitivity of ataxia-telangiectasia. , 1992, Radiation research.

[50]  W. L. Fangman,et al.  The localization of replication origins on ARS plasmids in S. cerevisiae , 1987, Cell.