Viral replication centers and the DNA damage response in JC virus-infected cells

ABSTRACT Infected cells recognize viral replication as a DNA damage stress and elicit the ataxia telangiectasia-mutated (ATM)/p53-mediated DNA damage response signal transduction pathway as part of the host surveillance mechanisms, which ultimately induces the irreversible cell cycle arrest and apoptosis. Viruses have evolved a variety of mechanisms to counteract this host intracellular innate immunity. Kaposi's sarcoma-associated herpesvirus (KSHV) viral interferon regulatory factor 1 (vIRF1) interacts with the cellular p53 tumor suppressor through its central DNA binding domain, and this interaction inhibits transcriptional activation of p53. Here, we further demonstrate that KSHV vIRF1 downregulates the total p53 protein level by facilitating its proteasome-mediated degradation. Detailed biochemical study showed that vIRF1 interacted with cellular ATM kinase through its carboxyl-terminal transactivation domain and that this interaction blocked the activation of ATM kinase activity induced by DNA damage stress. As a consequence, vIRF1 expression greatly reduced the level of serine 15 phosphorylation of p53, resulting in an increase of p53 ubiquitination and thereby a decrease of its protein stability. These results indicate that KSHV vIRF1 comprehensively compromises an ATM/p53-mediated DNA damage response checkpoint by targeting both upstream ATM kinase and downstream p53 tumor suppressor, which might circumvent host growth surveillance and facilitate viral replication in infected cells.

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SARS-CoV-2 triggers DNA damage response in Vero E6 cells

10.1101/2021.09.08.459535 ◽

2021 ◽

Author(s):

Joshua Victor ◽

Jamie Deutsch ◽

Annalis Whitaker ◽

Erica N. Lamkin ◽

Anthony March ◽

...

Keyword(s):

Dna Damage ◽

Dna Damage Response ◽

Host Cells ◽

Kidney Cells ◽

The Novel ◽

African Green Monkey Kidney ◽

Downstream Effector ◽

Damage Response ◽

Green Monkey ◽

Infected Cells

AbstractThe novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus responsible for the current COVID-19 pandemic and has now infected more than 200 million people with more than 4 million deaths globally. Recent data suggest that symptoms and general malaise may continue long after the infection has ended in recovered patients, suggesting that SARS-CoV-2 infection has profound consequences in the host cells. Here we report that SARS-CoV-2 infection can trigger a DNA damage response (DDR) in African green monkey kidney cells (Vero E6). We observed a transcriptional upregulation of the Ataxia telangiectasia and Rad3 related protein (ATR) in infected cells. In addition, we observed enhanced phosphorylation of CHK1, a downstream effector of the ATR DNA damage response, as well as H2AX. Strikingly, SARS-CoV-2 infection lowered the expression of TRF2 shelterin-protein complex, and reduced telomere lengths in infected Vero E6 cells. Thus, our observations suggest SARS-CoV-2 may have pathological consequences to host cells beyond evoking an immunopathogenic immune response.

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Regulation of NKG2D and DNAM-1 ligand expression in HCMV infected fibroblasts: role of the DNA damage response pathway and of viral replication.

Frontiers in Immunology ◽

10.3389/conf.fimmu.2013.02.00253 ◽

2013 ◽

Vol 4 ◽

Author(s):

Pignoloni Benedetta ◽

Dell'Oste Valentina ◽

Gribaudo Giorgio ◽

Cippitelli Marco ◽

Fionda Cinzia ◽

...

Keyword(s):

Dna Damage ◽

Viral Replication ◽

Dna Damage Response ◽

Damage Response ◽

Ligand Expression ◽

Dna Damage Response Pathway

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Viral DNA Replication-Dependent DNA Damage Response Activation during BK Polyomavirus Infection

Journal of Virology ◽

10.1128/jvi.03650-14 ◽

2015 ◽

Vol 89 (9) ◽

pp. 5032-5039 ◽

Cited By ~ 26

Author(s):

Brandy Verhalen ◽

Joshua L. Justice ◽

Michael J. Imperiale ◽

Mengxi Jiang

Keyword(s):

Bone Marrow ◽

Dna Damage ◽

Viral Replication ◽

Dna Damage Response ◽

Marrow Transplant ◽

Viral Dna ◽

Viral Dna Replication ◽

Transplant Patients ◽

Bk Polyomavirus ◽

Damage Response

ABSTRACTBK polyomavirus (BKPyV) reactivation is associated with severe human disease in kidney and bone marrow transplant patients. The interplay between viral and host factors that regulates the productive infection process remains poorly understood. We have previously reported that the cellular DNA damage response (DDR) is activated upon lytic BKPyV infection and that its activation is required for optimal viral replication in primary kidney epithelial cells. In this report, we set out to determine what viral components are responsible for activating the two major phosphatidylinositol 3-kinase-like kinases (PI3KKs) involved in the DDR: ataxia telangiectasia mutated (ATM) kinase and ATM and Rad3-related (ATR) kinase. Using a combination of UV treatment, lentivirus transduction, and mutant virus infection experiments, our results demonstrate that neither the input virus nor the expression of large T antigen (TAg) alone is sufficient to trigger the activation of ATM or ATR in our primary culture model. Instead, our data suggest that the activation of both the ATM- and ATR-mediated DDR pathways is linked to viral DNA replication. Intriguingly, a TAg mutant virus that is unable to activate the DDR causes substantial host DNA damage. Our study provides insight into how DDRs are activated by polyomaviruses in primary cells with intact cell cycle checkpoints and how the activation might be linked to the maintenance of host genome stability.IMPORTANCEPolyomaviruses are opportunistic pathogens that are associated with several human diseases under immunosuppressed conditions. BK polyomavirus (BKPyV) affects mostly kidney and bone marrow transplant patients. The detailed replication mechanism of these viruses remains to be determined. We have previously reported that BKPyV activates the host DNA damage response (DDR), a response normally used by the host cell to combat genotoxic stress, to aid its own replication. In this study, we identified that the trigger for DDR activation is viral replication. Furthermore, we show that the virus is able to cause host DNA damage in the absence of viral replication and DDR activation. These results suggest an intricate relationship between viral replication, DDR activation, and host genome instability.

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Differential Requirements of the C Terminus of Nbs1 in Suppressing Adenovirus DNA Replication and Promoting Concatemer Formation

Journal of Virology ◽

10.1128/jvi.00900-08 ◽

2008 ◽

Vol 82 (17) ◽

pp. 8362-8372 ◽

Cited By ~ 40

Author(s):

Seema S. Lakdawala ◽

Rachel A. Schwartz ◽

Kevin Ferenchak ◽

Christian T. Carson ◽

Brian P. McSharry ◽

...

Keyword(s):

Dna Damage ◽

Dna Replication ◽

Viral Replication ◽

Dna Damage Response ◽

Viral Genomes ◽

Mrn Complex ◽

E4 Region ◽

C Terminus ◽

Damage Response ◽

Cellular Dna

ABSTRACT Adenoviruses (Ad) with the early region E4 deleted (E4-deleted virus) are defective for DNA replication and late protein synthesis. Infection with E4-deleted viruses results in activation of a DNA damage response, accumulation of cellular repair factors in foci at viral replication centers, and joining together of viral genomes into concatemers. The cellular DNA repair complex composed of Mre11, Rad50, and Nbs1 (MRN) is required for concatemer formation and full activation of damage signaling through the protein kinases Ataxia-telangiectasia mutated (ATM) and ATM-Rad3-related (ATR). The E4orf3 and E4orf6 proteins expressed from the E4 region of Ad type 5 (Ad5) inactivate the MRN complex by degradation and mislocalization, and prevent the DNA damage response. Here we investigated individual contributions of the MRN complex, concatemer formation, and damage signaling to viral DNA replication during infection with E4-deleted virus. Using virus mutants, short hairpin RNA knockdown and hypomorphic cell lines, we show that inactivation of MRN results in increased viral replication. We demonstrate that defective replication in the absence of E4 is not due to concatemer formation or DNA damage signaling. The C terminus of Nbs1 is required for the inhibition of Ad DNA replication and recruitment of MRN to viral replication centers. We identified regions of Nbs1 that are differentially required for concatemer formation and inhibition of Ad DNA replication. These results demonstrate that targeting of the MRN complex explains the redundant functions of E4orf3 and E4orf6 in promoting Ad DNA replication. Understanding how MRN impacts the adenoviral life cycle will provide insights into the functions of this DNA damage sensor.

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Interaction of the Human Papillomavirus E1 Helicase with UAF1-USP1 Promotes Unidirectional Theta Replication of Viral Genomes

mBio ◽

10.1128/mbio.00152-19 ◽

2019 ◽

Vol 10 (2) ◽

Cited By ~ 1

Author(s):

Marit Orav ◽

David Gagnon ◽

Jacques Archambault

Keyword(s):

Dna Damage ◽

Dna Damage Response ◽

Fanconi Anemia ◽

Human Papillomaviruses ◽

Viral Genomes ◽

Damage Response ◽

Theta Replication ◽

Infected Cells ◽

Cellular Dna ◽

Medical Burden

ABSTRACTHuman papillomaviruses (HPVs) are important pathogens with a significant medical burden. HPV genomes replicate in infected cells via bidirectional theta replication and a poorly understood unidirectional mechanism. In this report, we provide evidence that the previously described interaction between the viral E1 helicase and the cellular UAF1-USP1 deubiquitinating enzyme complex, a member of the Fanconi anemia DNA damage response pathway, is required for the completion of the bidirectional theta replication of the HPV11 genome and the subsequent initiation of the unidirectional replication. We show that unidirectional replication proceeds via theta structures and is supported by the cellular Bloom helicase, which interacts directly with E1 and whose engagement in HPV11 replication requires UAF1-USP1 activity. We propose that the unidirectional replication of the HPV11 genome initiates from replication fork restart events. These findings suggest a new role for the Fanconi anemia pathway in HPV replication.IMPORTANCEHuman papillomaviruses (HPVs) are important pathogens that replicate their double-stranded circular DNA genome in the nucleus of infected cells. HPV genomes replicate in infected cells via bidirectional theta replication and a poorly understood unidirectional mechanism, and the onset of viral replication requires the engagement of cellular DNA damage response pathways. In this study, we showed that the previously described interaction between the viral E1 helicase and the cellular UAF1-USP1 complex is necessary for the completion of bidirectional replication and the subsequent initiation of the unidirectional replication mechanism. Our results suggest HPVs may use the cellular Fanconi anemia DNA damage pathway to achieve the separation of daughter molecules generated by bidirectional theta replication. Additionally, our results indicate that the unidirectional replication of the HPV genome is initiated from restarted bidirectional theta replication forks.

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Hijacking the DNA Damage Response to Enhance Viral Replication: γ-Herpesvirus 68 orf36 Phosphorylates Histone H2AX

Molecular Cell ◽

10.1016/j.molcel.2007.08.004 ◽

2007 ◽

Vol 27 (4) ◽

pp. 689

Author(s):

Anyong Xie ◽

Ralph Scully

Keyword(s):

Dna Damage ◽

Viral Replication ◽

Dna Damage Response ◽

Histone H2ax ◽

Damage Response

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