Evidence for the receipt of DNA damage stimuli by PML nuclear domains

Promyelocytic leukaemia nuclear domains (PML‐NDs) comprise a shell of PML protein and many labile cargo proteins. The nature of their cargo, their juxtaposition to foci of damaged DNA following ionizing radiation (IR), and the altered DNA damage responses in PML null cells all implicate PML‐NDs in the DNA damage response. In this work, the propensity of PML‐NDs to increase in number and decrease in size following IR has been studied. Serial quantitative studies of endogenous PML‐NDs prove that the PML‐ND response to IR is not the result of the asymmetry in cell cycle distribution that can follow IR, but reflects more directly the process of DNA damage. The response is swift, sensitive (evident after 1 Gy), and potentially reversible in untransformed fibroblasts. In these cells and in HCT116 colon cancer cells, failure to restore PML‐ND number within 24 h correlates with later loss of growth potential—in fibroblasts, through prolonged cell cycle arrest and in HCT116 cells, through apoptosis. Failure to express an intact ATM/CHK2 DNA damage signalling pathway in either cell type leads to a delay in the PML‐ND response to IR. Conversely, cell cycle progression following IR in cells that detect damaged DNA accelerates PML‐ND reorganization. Collectively, these data show that the increase in PML‐ND number seen after irradiation is, in part, triggered by the receipt of the DNA damage stimulus. The senescent cell state is also associated with chronic DNA damage and Hayflick‐limited fibroblasts were found to express nuclei with elevated numbers of PML‐NDs before IR that remained unresponsive to IR. Though the underlying reasons for damage‐induced PML alteration remain obscure, it is noteworthy that significant numbers of PML‐NDs juxtapose with ionizing radiation‐induced foci after IR. The co‐regulation of these structures may necessitate the stereotyped increases in PML‐ND number following damage. Copyright © 2007 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

[1]  A. Marson,et al.  PML bodies control the nuclear dynamics and function of the CHFR mitotic checkpoint protein , 2004, Nature Structural &Molecular Biology.

[2]  T. Ørntoft,et al.  DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis , 2005, Nature.

[3]  Q. Zhan,et al.  The mismatch repair system is required for S-phase checkpoint activation , 2003, Nature Genetics.

[4]  R. Muschel,et al.  DNA Damage in HeLa Cells Induced Arrest at a Discrete Point in G2 Phase as Defined by CENP-F Localization , 2003, Radiation research.

[5]  Nicholas J. McGlincy,et al.  Isoforms of the promyelocytic leukemia protein differ in their effects on ND10 organization. , 2005, Experimental cell research.

[6]  G. Dellaire,et al.  Chromatin Contributes to Structural Integrity of Promyelocytic Leukemia Bodies through a SUMO-1-independent Mechanism* , 2004, Journal of Biological Chemistry.

[7]  P. Pandolfi,et al.  Pml is essential for multiple apoptotic pathways , 1998, Nature Genetics.

[8]  C. Prives,et al.  The Chk2 protein kinase. , 2004, DNA repair.

[9]  R. van Driel,et al.  Cell cycle regulation of PML modification and ND10 composition. , 1999, Journal of cell science.

[10]  E. Bowman,et al.  UV-C-induced DNA damage leads to p53-dependent nuclear trafficking of PML , 2003, Oncogene.

[11]  J. Bisi,et al.  PML-dependent apoptosis after DNA damage is regulated by the checkpoint kinase hCds1/Chk2 , 2002, Nature Cell Biology.

[12]  H. de Thé,et al.  Characterization of endogenous human promyelocytic leukemia isoforms. , 2006, Cancer research.

[13]  S. Minucci,et al.  PML NBs associate with the hMre11 complex and p53 at sites of irradiation induced DNA damage , 2002, Oncogene.

[14]  Dimitris Kletsas,et al.  Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions , 2005, Nature.

[15]  J. Barrett,et al.  Senescing human cells and ageing mice accumulate DNA lesions with unrepairable double-strand breaks , 2004, Nature Cell Biology.

[16]  Sara L. Tuttle,et al.  The promyelocytic leukemia protein PML regulates c-Jun function in response to DNA damage. , 2005, Blood.

[17]  Xiaolu Yang,et al.  Association of caspase-2 with the promyelocytic leukemia protein nuclear bodies , 2005, Cancer biology & therapy.

[18]  G. Dellaire,et al.  The number of PML nuclear bodies increases in early S phase by a fission mechanism , 2006, Journal of Cell Science.

[19]  R. Everett,et al.  ND10 Components Relocate to Sites Associated with Herpes Simplex Virus Type 1 Nucleoprotein Complexes during Virus Infection , 2005, Journal of Virology.

[20]  A. Wyllie,et al.  Stress responses of PML nuclear domains are ablated by ataxin‐1 and other nucleoprotein inclusions , 2004, The Journal of pathology.

[21]  K. Borden,et al.  Pondering the Promyelocytic Leukemia Protein (PML) Puzzle: Possible Functions for PML Nuclear Bodies , 2002, Molecular and Cellular Biology.

[22]  P. Lønning,et al.  Promyelocytic leukemia nuclear bodies are predetermined processing sites for damaged DNA , 2006, Journal of Cell Science.

[23]  Zhi-xiang Xu,et al.  PML Colocalizes with and Stabilizes the DNA Damage Response Protein TopBP1 , 2003, Molecular and Cellular Biology.

[24]  P. Pandolfi,et al.  Promyelocytic Leukemia Activates Chk2 by Mediating Chk2 Autophosphorylation* , 2006, Journal of Biological Chemistry.

[25]  Wei-Wei Zhang,et al.  Cell-cycle regulation of DNA damage-induced expression of the suppressor gene PML. , 1997, Biochemical and biophysical research communications.

[26]  Bert Vogelstein,et al.  Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21 , 1996, Nature.

[27]  Joe S. Mymryk,et al.  Size, position and dynamic behavior of PML nuclear bodies following cell stress as a paradigm for supramolecular trafficking and assembly , 2003, Journal of Cell Science.

[28]  K. Kohn,et al.  UCN-01 inhibits p53 up-regulation and abrogates gamma-radiation-induced G(2)-M checkpoint independently of p53 by targeting both of the checkpoint kinases, Chk2 and Chk1. , 2002, Cancer research.

[29]  H. Tanke,et al.  Mobile foci of Sp100 do not contain PML: PML bodies are immobile but PML and Sp100 proteins are not. , 2002, Journal of structural biology.

[30]  P. Pandolfi,et al.  Role of PML and the PML-nuclear body in the control of programmed cell death , 2003, Oncogene.

[31]  L. Latonen,et al.  Cellular stress and DNA damage invoke temporally distinct Mdm2, p53 and PML complexes and damage-specific nuclear relocalization , 2003, Journal of Cell Science.

[32]  Graham Dellaire,et al.  PML nuclear bodies: dynamic sensors of DNA damage and cellular stress , 2004, BioEssays : news and reviews in molecular, cellular and developmental biology.