PML nuclear bodies are recruited to persistent DNA damage lesions in an RNF168-53BP1 dependent manner and contribute to DNA repair

AbstractMaintaining genomic stability is inevitable for organism survival and it is challenged by mutagenic agents, which include ultraviolet (UV) radiation. Whenever DNA damage occurs, it is sensed by DNA-repairing proteins and thereby performing the DNA-repair mechanism. Specifically, in response to DNA damage, H2AX is a key protein involved in initiating the DNA-repair processes. In this present study, we investigate the effect of UV-C on earthworm, Perionyx excavatus and analyzed the DNA-damage response. Briefly, we expose the worms to different doses of UV-C and find that worms are highly sensitive to UV-C. As a primary response, earthworms produce coelomic fluid followed by autotomy. However, tissue inflammation followed by death is observed when we expose worm to increased doses of UV-C. In particular, UV-C promotes damages in skin layers and on the contrary, it mediates the chloragogen and epithelial outgrowth in intestinal tissues. Furthermore, UV-C promotes DNA damages followed by upregulation of H2AX on dose-dependent manner. Our finding confirms DNA damage caused by UV-C is directly proportional to the expression of H2AX. In short, we conclude that H2AX is present in the invertebrate earthworm, which plays an evolutionarily conserved role in DNA damage event as like that in higher animals.

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Yeast Rpn4 Links the Proteasome and DNA Repair via RAD52 Regulation

International Journal of Molecular Sciences ◽

10.3390/ijms21218097 ◽

2020 ◽

Vol 21 (21) ◽

pp. 8097

Author(s):

Daria S. Spasskaya ◽

Nonna I. Nadolinskaia ◽

Vera V. Tutyaeva ◽

Yuriy P. Lysov ◽

Vadim L. Karpov ◽

...

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Proteasome Inhibition ◽

26S Proteasome ◽

Dependent Manner ◽

Dna Repair Genes ◽

Damage Resistance ◽

Dna Damaging Agents ◽

Mutant Strains ◽

Repair Genes

Environmental and intracellular factors often damage DNA, but multiple DNA repair pathways maintain genome integrity. In yeast, the 26S proteasome and its transcriptional regulator and substrate Rpn4 are involved in DNA damage resistance. Paradoxically, while proteasome dysfunction may induce hyper-resistance to DNA-damaging agents, Rpn4 malfunction sensitizes yeasts to these agents. Previously, we proposed that proteasome inhibition causes Rpn4 stabilization followed by the upregulation of Rpn4-dependent DNA repair genes and pathways. Here, we aimed to elucidate the key Rpn4 targets responsible for DNA damage hyper-resistance in proteasome mutants. We impaired the Rpn4-mediated regulation of candidate genes using the CRISPR/Cas9 system and tested the sensitivity of mutant strains to 4-NQO, MMS and zeocin. We found that the separate or simultaneous deregulation of 19S or 20S proteasome subcomplexes induced MAG1, DDI1, RAD23 and RAD52 in an Rpn4-dependent manner. Deregulation of RAD23, DDI1 and RAD52 sensitized yeast to DNA damage. Genetic, epigenetic or dihydrocoumarin-mediated RAD52 repression restored the sensitivity of the proteasome mutants to DNA damage. Our results suggest that the Rpn4-mediated overexpression of DNA repair genes, especially RAD52, defines the DNA damage hyper-resistant phenotype of proteasome mutants. The developed yeast model is useful for characterizing drugs that reverse the DNA damage hyper-resistance phenotypes of cancers.

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Arsenic-Induced SUMO-Dependent Recruitment of RNF4 into PML Nuclear Bodies

Molecular Biology of the Cell ◽

10.1091/mbc.e10-05-0449 ◽

2010 ◽

Vol 21 (23) ◽

pp. 4227-4239 ◽

Cited By ~ 41

Author(s):

Marie-Claude Geoffroy ◽

Ellis G. Jaffray ◽

Katherine J. Walker ◽

Ronald T. Hay

Keyword(s):

Retinoic Acid Receptor ◽

E3 Ligase ◽

Nuclear Body ◽

Promyelocytic Leukemia ◽

Nuclear Bodies ◽

Dependent Manner ◽

Nuclear Trafficking ◽

Sumo Modification ◽

Ubiquitin E3 Ligase ◽

Pml Nuclear Bodies

In acute promyelocytic leukemia (APL), the promyelocytic leukemia (PML) protein is fused to the retinoic acid receptor alpha (RAR). Arsenic is an effective treatment for this disease as it induces SUMO-dependent ubiquitin-mediated proteasomal degradation of the PML-RAR fusion protein. Here we analyze the nuclear trafficking dynamics of PML and its SUMO-dependent ubiquitin E3 ligase, RNF4 in response to arsenic. After administration of arsenic, PML immediately transits into nuclear bodies where it undergoes SUMO modification. This initial recruitment of PML into nuclear bodies is not dependent on RNF4, but RNF4 quickly follows PML into the nuclear bodies where it is responsible for ubiquitylation of SUMO-modified PML and its degradation by the proteasome. While arsenic restricts the mobility of PML, FRAP analysis indicates that RNF4 continues to rapidly shuttle into PML nuclear bodies in a SUMO-dependent manner. Under these conditions FRET studies indicate that RNF4 interacts with SUMO in PML bodies but not directly with PML. These studies indicate that arsenic induces the rapid reorganization of the cell nucleus by SUMO modification of nuclear body-associated PML and uptake of the ubiquitin E3 ligase RNF4 leading to the ubiquitin-mediated degradation of PML.

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Transcriptional Regulation Is Affected by Subnuclear Targeting of Reporter Plasmids to PML Nuclear Bodies

Molecular and Cellular Biology ◽

10.1128/mcb.00636-06 ◽

2006 ◽

Vol 26 (23) ◽

pp. 8814-8825 ◽

Cited By ~ 26

Author(s):

Gregory J. Block ◽

Christopher H. Eskiw ◽

Graham Dellaire ◽

David P. Bazett-Jones

Keyword(s):

Transcriptional Regulation ◽

Transcriptional Activation ◽

Plasmid Dna ◽

Simian Virus 40 ◽

Simian Virus ◽

Nuclear Bodies ◽

Luciferase Reporter ◽

Dependent Manner ◽

Reporter Plasmid ◽

Pml Nuclear Bodies

ABSTRACT Whereas the PML protein has been reported to have both transcriptional coactivator and corepressor potential, the contribution of the PML nuclear body (PML NB) itself to transcriptional regulation is not well understood. Here we demonstrate that plasmid DNA artificially tethered to PML or the PML NB-targeting domain of Sp100 is preferentially localized to PML NBs. Using the tethering technique, we targeted a simian virus 40 promoter-driven luciferase reporter plasmid to PML NBs, resulting in the repression of the transgene transcriptional activity. Conversely, the tethering of a cytomegalovirus promoter-containing reporter plasmid resulted in activation. Targeting a minimal eukaryotic promoter did not affect its activity. The expression of targeted promoters could be modulated by altering the cellular concentration of PML NB components, including Sp100 and isoforms of the PML protein. Finally, we demonstrate that ICP0, the promiscuous herpes simplex virus transactivator, increases the level of transcriptional activation of plasmid DNA tethered to the PML NB. We conclude that when PML NB components are artificially tethered to reporter plasmids, the PML NB contributes to the regulation of the tethered DNA in a promoter-dependent manner. Our findings demonstrate that transient transcription assays are sensitive to the subnuclear localization of the transgene plasmid.

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The Disruption of ND10 during Herpes Simplex Virus Infection Correlates with the Vmw110- and Proteasome-Dependent Loss of Several PML Isoforms

Journal of Virology ◽

10.1128/jvi.72.8.6581-6591.1998 ◽

1998 ◽

Vol 72 (8) ◽

pp. 6581-6591 ◽

Cited By ~ 298

Author(s):

Roger D. Everett ◽

Paul Freemont ◽

Hisato Saitoh ◽

Mary Dasso ◽

Anne Orr ◽

...

Keyword(s):

Virus Infection ◽

Regulatory Protein ◽

Biochemical Properties ◽

Nuclear Bodies ◽

Dependent Manner ◽

Pml Nuclear Bodies ◽

Protease Enzyme ◽

Specific Protease ◽

Nuclear Structures ◽

Simplex Virus

ABSTRACT The small nuclear structures known as ND10 or PML nuclear bodies have been implicated in a variety of cellular processes including response to stress and interferons, oncogenesis, and viral infection, but little is known about their biochemical properties. Recently, a ubiquitin-specific protease enzyme (named HAUSP) and a ubiquitin-homology family protein (PIC1) have been found associated with ND10. HAUSP binds strongly to Vmw110, a herpesvirus regulatory protein which has the ability to disrupt ND10, while PIC1 was identified as a protein which interacts with PML, the prototype ND10 protein. We have investigated the role of ubiquitin-related pathways in the mechanism of ND10 disruption by Vmw110 and the effect of virus infection on PML stability. The results show that the disruption of ND10 during virus infection correlates with the loss of several PML isoforms and this process is dependent on active proteasomes. The PML isoforms that are most sensitive to virus infection correspond closely to those which have recently been identified as being covalently conjugated to PIC1. In addition, a large number of PIC1-protein conjugates can be detected following transfection of a PIC1 expression plasmid, and many of these are also eliminated in a Vmw110-dependent manner during virus infection. These observations provide a biochemical mechanism to explain the observed effects of Vmw110 on ND10 and suggest a simple yet powerful mechanism by which Vmw110 might function during virus infection.

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ATM controls DNA repair and mitochondria transfer between neighboring cells

Cell Communication and Signaling ◽

10.1186/s12964-019-0472-x ◽

2019 ◽

Vol 17 (1) ◽

Cited By ~ 2

Author(s):

Sha Jin ◽

Nils Cordes

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Dependent Manner ◽

Intercellular Signaling ◽

Tissue Integrity ◽

Damage Repair ◽

Repair Protein ◽

Mitochondria Transfer ◽

Dna Repair Protein ◽

Direct Cell

Abstract Intercellular communication is essential for multicellular tissue vitality and homeostasis. We show that healthy cells message protective signals through direct cell–cell connections to adjacent DNA–damaged cells in a microtubule–dependent manner. In DNA–damaged cells, mitochondria restoration is facilitated by fusion with undamaged mitochondria from healthy cells and their DNA damage repair is optimized in presence of healthy cells. Both, mitochondria transfer and intercellular signaling for an enhanced DNA damage response are critically regulated by the activity of the DNA repair protein ataxia telangiectasia mutated (ATM). These healthy–to–damaged prosurvival processes sustain normal tissue integrity and may be exploitable for overcoming resistance to therapy in diseases such as cancer.

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