Replication Stress and Mitotic Dysfunction in Cells Expressing Simian Virus 40 Large T Antigen

Hu, Liang; Filippakis, Harilaos; Huang, Hongbiao; Yen, Tim J.; Gjoerup, Ole

doi:10.1128/jvi.02224-13

Cited by 18 publications

(15 citation statements)

References 91 publications

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“…This suggests that BKPyV is capable of causing host damage, possibly through TAg, but that this damage is likely repaired by the activated DDRs during normal replication. Alternatively, it is possible that BKPyV infection results in host replication stress and mitotic dysfunction, similar to what has been reported for SV40 TAg (32). During normal infection, however, the damage is not apparent because the cells are arrested in G 2 (14).…”

Section: Discussionmentioning

confidence: 74%

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

Verhalen

Justice

Imperiale

et al. 2015

J Virol

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BK 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. The BK polyomavirus (BKPyV) is a ubiquitous opportunistic human pathogen which causes severe disease in immunocompromised patients (1). BKPyV is thought to be acquired through the respiratory route during early childhood, and by adulthood, up to 90% of the general population becomes seropositive (2). Following primary exposure, the virus establishes a lifelong, subclinical, persistent infection in the genitourinary tract. BKPyV can reactivate from the persistent state under immunosuppressed conditions, most commonly in kidney transplant patients, resulting in viral shedding in urine or blood and, ultimately, polyomavirus-associated nephropathy, a significant cause of renal dysfunction (3). There are no FDA-approved therapies for BKPyV infection, and the usual treatment is to reduce immu...

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Section: Discussionmentioning

confidence: 74%

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

Verhalen

Justice

Imperiale

et al. 2015

J Virol

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“…Transgenic mice in which cell cycle reactivation in neurons is forced by SV40 T antigen via the tet-on/off system show signs of mitotic re-entry (e.g., PCNA, cyclin B1, MPM2) and Aβ deposits and phosphorylated tau in the brain (Park, Hallows, Chakrabarty, Davies, & Vincent, 2007). Expression of SV40 T antigen causes replication stress, mitotic dysfunction, and aneuploidy (Hu, Filippakis, Huang, Yen, & Gjoerup, 2013), suggesting a link among mitotic re-entry, aneuploidy, and AD pathology.…”

Section: Forced Cell Cycle Re-entry Resulted In Amyloidbeta and P-tmentioning

confidence: 99%

“Amyloid‐beta accumulation cycle” as a prevention and/or therapy target for Alzheimer's disease

et al. 2020

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The cell cycle and its regulators are validated targets for cancer drugs. Reagents that target cells in a specific cell cycle phase (e.g., antimitotics or DNA synthesis inhibitors/replication stress inducers) have demonstrated success as broad‐spectrum anticancer drugs. Cyclin‐dependent kinases (CDKs) are drivers of cell cycle transitions. A CDK inhibitor, flavopiridol/alvocidib, is an FDA‐approved drug for acute myeloid leukemia. Alzheimer's disease (AD) is another serious issue in contemporary medicine. The cause of AD remains elusive, although a critical role of latent amyloid‐beta accumulation has emerged. Existing AD drug research and development targets include amyloid, amyloid metabolism/catabolism, tau, inflammation, cholesterol, the cholinergic system, and other neurotransmitters. However, none have been validated as therapeutically effective targets. Recent reports from AD‐omics and preclinical animal models provided data supporting the long‐standing notion that cell cycle progression and/or mitosis may be a valid target for AD prevention and/or therapy. This review will summarize the recent developments in AD research: (a) Mitotic re‐entry, leading to the “amyloid‐beta accumulation cycle,” may be a prerequisite for amyloid‐beta accumulation and AD pathology development; (b) AD‐associated pathogens can cause cell cycle errors; (c) thirteen among 37 human AD genetic risk genes may be functionally involved in the cell cycle and/or mitosis; and (d) preclinical AD mouse models treated with CDK inhibitor showed improvements in cognitive/behavioral symptoms. If the “amyloid‐beta accumulation cycle is an AD drug target” concept is proven, repurposing of cancer drugs may emerge as a new, fast‐track approach for AD management in the clinic setting.

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“…It is also supported by our DNase treatment data, which suggest that the defective genomes were encapsidated. Perhaps generation of defective genomes was driven by constitutive SV40 LTag expression, as this protein is known to cause replicative stress and mitotic dysfunction leading to both structural and numerical chromosome instability (63). Whatever the mechanism, generation of defective mutants in polyomavirus cell culture has previously been observed and seems to depend on both host and viral factors (64,65).…”

Section: Discussionmentioning

confidence: 99%

The Human Fetal Glial Cell Line SVG p12 Contains Infectious BK Polyomavirus

Henriksen

Tylden

Dumoulin

et al. 2014

J Virol

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The human fetal glial cell line SVG was generated in 1985 by transfecting primary fetal brain cells with a plasmid containing an origin-defective mutant of simian virus 40 (SV40). The cells, which express SV40 large T-antigen, support the replication of human JC polyomavirus (JCPyV) and have been used for JCPyV studies but also for other studies in which cells of neural origin were desirable. We intended to use the SVG p12 cells from ATCC for antiviral drug studies with JCPyV. However, during initial experiments, immunofluorescence microscopy controls unexpectedly revealed cells expressing the late viral proteins VP1, VP2/ VP3, and agno. This was confirmed by Western blotting. Since our agnoprotein antiserum is specific for BKPyV agnoprotein, infection with BKPyV was suspected. Indeed, specific BKPyV PCR of SVG p12 supernatants revealed a viral load of >1 ؋ 10

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Replication Stress and Mitotic Dysfunction in Cells Expressing Simian Virus 40 Large T Antigen

Cited by 18 publications

References 91 publications

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

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

“Amyloid‐beta accumulation cycle” as a prevention and/or therapy target for Alzheimer's disease

The Human Fetal Glial Cell Line SVG p12 Contains Infectious BK Polyomavirus

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