Open Access

Microorganisms, volume 7, issue 11, pages 533

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

Brezgin Sergey ^{1, 2}

Kostyusheva Anastasiia ²

Bayurova Ekaterina ^{3, 4}

Gordeychuk Ilya ^{3, 4, 5}

Isaguliants Maria ^{3, 4, 6, 7}

Goptar Irina ⁸

Nikiforova Anastasiia ⁸

Smirnov Valery ¹

Volchkova Elena ⁵

Glebe Dieter ⁹

Kostyushev Dmitry ²

Chulanov Vladimir ^{2, 5, 10}

Hide authors affiliations Show authors affiliations: 10 affiliations

Institute of Immunology, Federal Medical Biological Agency, 115522 Moscow, Russia |

National Medical Research Center for Tuberculosis and Infectious Diseases, 127994 Moscow, Russia |

Chumakov Federal Scientific Center for Research and Development of Immune and Biological Products of Russian Academy of Sciences, 108819 Moscow, Russia |

⁴

NF Gamaleya Research Center of Epidemiology and Microbiology, 123098 Moscow, Russia |

⁵

Sechenov First Moscow State Medical University, 119146 Moscow, Russia |

⁶

Karolinska Institutet, SE-171 76 Stockholm, Sweden |

⁷

Riga Stradins University, LV-1007 Riga, Latvia |

⁸

Izmerov Research Institute of Occupational Health, 105275 Moscow, Russia |

⁹

Institute of Medical Virology, University of Giessen, 35392 Giessen, Germany |

¹⁰

Central Research Institute of Epidemiology, 111123 Moscow, Russia |

Publication type: Journal Article

Publication date: 2019-11-06

Multidisciplinary Digital Publishing Institute (MDPI)

Journal: Microorganisms

Quartile SCImago

Quartile WOS

Impact factor: 4.5

ISSN: 20762607

DOI: 10.3390/microorganisms7110533

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PubMed ID: 31698767

Microbiology (medical)

Microbiology

Virology

Abstract

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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Citations by journals

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International Journal of Molecular Sciences	International Journal of Molecular Sciences, 2, 20% International Journal of Molecular Sciences 2 publications, 20%
Viruses	Viruses, 2, 20% Viruses 2 publications, 20%
Vaccines	Vaccines, 1, 10% Vaccines 1 publication, 10%
World Journal of Virology	World Journal of Virology, 1, 10% World Journal of Virology 1 publication, 10%
Antiviral Research	Antiviral Research, 1, 10% Antiviral Research 1 publication, 10%
Journal of Medical Virology	Journal of Medical Virology, 1, 10% Journal of Medical Virology 1 publication, 10%
Virus Evolution	Virus Evolution, 1, 10% Virus Evolution 1 publication, 10%
Expert Reviews in Molecular Medicine	Expert Reviews in Molecular Medicine, 1, 10% Expert Reviews in Molecular Medicine 1 publication, 10%
	1 2

Citations by publishers

	1 2 3 4 5
Multidisciplinary Digital Publishing Institute (MDPI)	Multidisciplinary Digital Publishing Institute (MDPI), 5, 50% Multidisciplinary Digital Publishing Institute (MDPI) 5 publications, 50%
Baishideng Publishing Group	Baishideng Publishing Group, 1, 10% Baishideng Publishing Group 1 publication, 10%
Elsevier	Elsevier, 1, 10% Elsevier 1 publication, 10%
Wiley	Wiley, 1, 10% Wiley 1 publication, 10%
Oxford University Press	Oxford University Press, 1, 10% Oxford University Press 1 publication, 10%
Cambridge University Press	Cambridge University Press, 1, 10% Cambridge University Press 1 publication, 10%
	1 2 3 4 5

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Brezgin S. et al. Replenishment of Hepatitis B Virus cccDNA Pool Is Restricted by Baseline Expression of Host Restriction Factors In Vitro // Microorganisms. 2019. Vol. 7. No. 11. p. 533.

GOST all authors (up to 50) Copy

Brezgin S., Kostyusheva A., Bayurova E., Gordeychuk I., Isaguliants M., Goptar I., Nikiforova A., Smirnov V., Volchkova E., Glebe D., Kostyushev D., Chulanov V. Replenishment of Hepatitis B Virus cccDNA Pool Is Restricted by Baseline Expression of Host Restriction Factors In Vitro // Microorganisms. 2019. Vol. 7. No. 11. p. 533.

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RIS Copy

TY - JOUR

DO - 10.3390/microorganisms7110533

UR - https://doi.org/10.3390%2Fmicroorganisms7110533

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

T2 - Microorganisms

AU - Kostyusheva, Anastasiia

AU - Goptar, Irina

AU - Kostyushev, Dmitry

AU - Smirnov, Valery

AU - Brezgin, Sergey

AU - Isaguliants, Maria

AU - Volchkova, Elena

AU - Glebe, Dieter

AU - Bayurova, Ekaterina

AU - Gordeychuk, Ilya

AU - Nikiforova, Anastasiia

AU - Chulanov, Vladimir

PY - 2019

DA - 2019/11/06 00:00:00

PB - Multidisciplinary Digital Publishing Institute (MDPI)

SP - 533

IS - 11

VL - 7

PMID - 31698767

SN - 2076-2607

ER -

BibTex |

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@article{2019_Brezgin,

author = {Anastasiia Kostyusheva and Irina Goptar and Dmitry Kostyushev and Valery Smirnov and Sergey Brezgin and Maria Isaguliants and Elena Volchkova and Dieter Glebe and Ekaterina Bayurova and Ilya Gordeychuk and Anastasiia Nikiforova and Vladimir Chulanov},

title = {Replenishment of Hepatitis B Virus cccDNA Pool Is Restricted by Baseline Expression of Host Restriction Factors In Vitro},

journal = {Microorganisms},

year = {2019},

volume = {7},

publisher = {Multidisciplinary Digital Publishing Institute (MDPI)},

month = {nov},

url = {https://doi.org/10.3390%2Fmicroorganisms7110533},

number = {11},

pages = {533},

doi = {10.3390/microorganisms7110533}

}

MLA

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Brezgin, Sergey, et al. “Replenishment of Hepatitis B Virus cccDNA Pool Is Restricted by Baseline Expression of Host Restriction Factors In Vitro.” Microorganisms, vol. 7, no. 11, Nov. 2019, p. 533. https://doi.org/10.3390%2Fmicroorganisms7110533.

Found error?

Publisher

Multidisciplinary Digital Publishing Institute (MDPI)

Journal

Microorganisms

Quartile SCImago

Quartile WOS

Impact factor

4.5

ISSN

20762607 (Electronic)

Labs

Laboratory of Genetic Technologies

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