ATM and ATR Expression Potentiates HBV Replication and Contributes to Reactivation of HBV Infection upon DNA Damage

Kostyusheva, Anastasiya; Brezgin, Sergey; Bayurova, Ekaterina; Gordeychuk, Ilya; Isaguliants, Maria; Goptar, Irina A.; Urusov, F. A.; Nikiforova, A. V.; Volchkova, E. V; Kostyushev, Dmitry; Chulanov, Vladimir

doi:10.3390/v11110997

Cited by 21 publications

(16 citation statements)

References 63 publications

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“…1 B ). Our test panel represented genes that were either previously implicated in host cell response to viral infections such as BRCA1, RAD51, ATM, ATR, PRKDC, MRE11 [ 22 , 23 ], or picked as a logical extension of the key genes in the DNA repair pathways, such as PARP1 , XPA, and CHK1 . Transcript and western blot analyses showed an activation of the ATR DNA damage response post SARS-CoV-2 infection at 48 hours.…”

Section: Resultsmentioning

confidence: 99%

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

Victor

Deutsch

Whitaker

et al. 2021

Biochemical and Biophysical Research Communications

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The 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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Section: Resultsmentioning

confidence: 99%

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

Victor

Deutsch

Whitaker

et al. 2021

Biochemical and Biophysical Research Communications

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“…1B). Our test panel represented genes that were either previously implicated in host cell response to viral infections such as BRCA1, RAD51, ATM, ATR, PRKDC, MRE11 21,22 , or picked as a logical extension of the key genes in the DNA repair pathways, such as PARP1, XPA, and CHK1.…”

Section: Resultsmentioning

confidence: 99%

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

Victor

Deutsch

Whitaker

et al. 2021

Preprint

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The 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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“…Constitutive expression of DNMT3A reduced levels of DNA-PKcs , RAD51 , and ATR in both cell lines (Figure 6C,D). Notably, DNMT3A may reduce HBV replication not only epigenetically, but also by diminishing ATM/ATR levels, as this axis is involved in HBV replication [17,43,44,45].…”

Section: Resultsmentioning

confidence: 99%

Replenishment of Hepatitis B Virus cccDNA Pool Is Restricted by Baseline Expression of Host Restriction Factors In Vitro

Brezgin

Kostyusheva²,

Bayurova

et al. 2019

Microorganisms

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Background: Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is the major cause of viral persistence in patients with chronic HBV infection. Understanding the mechanisms underlying stability and persistence of HBV cccDNA in hepatocytes is critical for developing novel therapeutics and managing chronic hepatitis B. In this study, we observed an unexpected increase in HBV cccDNA levels upon suppression of transcription by de novo DNA methyltransferase DNMT3A and uncovered additional mechanisms potentially involved in HBV cccDNA maintenance. Methods: HBV-expressing cell lines were transfected with a DNMT3A-expressing plasmid. Real-time PCR and HBsAg assays were used to assess the HBV replication rate. Cell cycling was analyzed by fluorescent cell sorting. CRISPR/Cas9 was utilized to abrogate expression of APOBEC3A and APOBEC3B. Alterations in the expression of target genes were measured by real-time PCR. Results: Similar to previous studies, HBV replication induced DNMT3A expression, which in turn, led to reduced HBV transcription but elevated HBV cccDNA levels (4- to 6-fold increase). Increased levels of HBV cccDNA were not related to cell cycling, as DNMT3A accelerated proliferation of infected cells and could not contribute to HBV cccDNA expansion by arresting cells in a quiescent state. At the same time, DNMT3A suppressed transcription of innate immunity factors including cytidine deaminases APOBEC3A and APOBEC3B. CRISPR/Cas9-mediated silencing of APOBEC3A and APOBEC3B transcription had minor effects on HBV transcription, but significantly increased HBV cccDNA levels, similar to DNMT3A. In an attempt to further analyze the detrimental effects of HBV and DNMT3A on infected cells, we visualized γ-H2AX foci and demonstrated that HBV inflicts and DNMT3A aggravates DNA damage, possibly by downregulating DNA damage response factors. Additionally, suppression of HBV replication by DNMT3A may be related to reduced ATM/ATR expression. Conclusion: Formation and maintenance of HBV cccDNA pools may be partially suppressed by the baseline expression of host inhibitory factors including APOBEC3A and APOBEC3B. HBV inflicts DNA damage both directly and by inducing DNMT3A expression.

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ATM and ATR Expression Potentiates HBV Replication and Contributes to Reactivation of HBV Infection upon DNA Damage

Cited by 21 publications

References 63 publications

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

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

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

Replenishment of Hepatitis B Virus cccDNA Pool Is Restricted by Baseline Expression of Host Restriction Factors In Vitro

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