Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair?

Xiuji Cui, Rebecca McAllister, Rajeev Boregowda, Ji A. Sohn, Felipe Cortes Ledesma, Keith W. Caldecott, Christoph Seeger, Jianming Hu

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Hepatitis B virus (HBV) replication and persistence are sustained by a nuclear episome, the covalently closed circular (CCC) DNA, which serves as the transcriptional template for all viral RNAs. CCC DNA is converted from a relaxed circular (RC) DNA in the virion early during infection as well as from RC DNA in intracellular progeny nucleocapsids via an intracellular amplification pathway. Current antiviral therapies suppress viral replication but cannot eliminate CCC DNA. Thus, persistence of CCC DNA remains an obstacle toward curing chronic HBV infection. Unfortunately, very little is known about how CCC DNA is formed. CCC DNA formation requires removal of the virally encoded reverse transcriptase (RT) protein from the 5' end of the minus strand of RC DNA. Tyrosyl DNA phosphodiesterase-2 (Tdp2) was recently identified as the enzyme responsible for cleavage of tyrosyl-5' DNA linkages formed between topoisomerase II and cellular DNA. Because the RT-DNA linkage is also a 5' DNA-phosphotyrosyl bond, it has been hypothesized that Tdp2 might be one of several elusive host factors required for CCC DNA formation. Therefore, we examined the role of Tdp2 in RC DNA deproteination and CCC DNA formation. We demonstrated Tdp2 can cleave the tyrosyl-minus strand DNA linkage using authentic HBV RC DNA isolated from nucleocapsids and using RT covalently linked to short minus strand DNA produced in vitro. On the other hand, our results showed that Tdp2 gene knockout did not block CCC DNA formation during HBV infection of permissive human hepatoma cells and did not prevent intracellular amplification of duck hepatitis B virus CCC DNA. These results indicate that although Tdp2 can remove the RT covalently linked to the 5' end of the HBV minus strand DNA in vitro, this protein might not be required for CCC DNA formation in vivo.

Original languageEnglish (US)
Article numbere0128401
JournalPloS one
Volume10
Issue number6
DOIs
StatePublished - Jun 16 2015

Fingerprint

circular DNA
Circular DNA
Hepatitis B virus
Viruses
Repair
Genes
Genome
genome
DNA
RNA-directed DNA polymerase
RNA-Directed DNA Polymerase
linkage (genetics)
Nucleocapsid
nucleocapsid
tyrosyl-DNA phosphodiesterase
Virus Diseases
virus replication
Amplification
Duck hepatitis B virus
Duck Hepatitis B Viruses

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Cui, X., McAllister, R., Boregowda, R., Sohn, J. A., Ledesma, F. C., Caldecott, K. W., ... Hu, J. (2015). Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair? PloS one, 10(6), [e0128401]. https://doi.org/10.1371/journal.pone.0128401
Cui, Xiuji ; McAllister, Rebecca ; Boregowda, Rajeev ; Sohn, Ji A. ; Ledesma, Felipe Cortes ; Caldecott, Keith W. ; Seeger, Christoph ; Hu, Jianming. / Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair?. In: PloS one. 2015 ; Vol. 10, No. 6.
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abstract = "Hepatitis B virus (HBV) replication and persistence are sustained by a nuclear episome, the covalently closed circular (CCC) DNA, which serves as the transcriptional template for all viral RNAs. CCC DNA is converted from a relaxed circular (RC) DNA in the virion early during infection as well as from RC DNA in intracellular progeny nucleocapsids via an intracellular amplification pathway. Current antiviral therapies suppress viral replication but cannot eliminate CCC DNA. Thus, persistence of CCC DNA remains an obstacle toward curing chronic HBV infection. Unfortunately, very little is known about how CCC DNA is formed. CCC DNA formation requires removal of the virally encoded reverse transcriptase (RT) protein from the 5' end of the minus strand of RC DNA. Tyrosyl DNA phosphodiesterase-2 (Tdp2) was recently identified as the enzyme responsible for cleavage of tyrosyl-5' DNA linkages formed between topoisomerase II and cellular DNA. Because the RT-DNA linkage is also a 5' DNA-phosphotyrosyl bond, it has been hypothesized that Tdp2 might be one of several elusive host factors required for CCC DNA formation. Therefore, we examined the role of Tdp2 in RC DNA deproteination and CCC DNA formation. We demonstrated Tdp2 can cleave the tyrosyl-minus strand DNA linkage using authentic HBV RC DNA isolated from nucleocapsids and using RT covalently linked to short minus strand DNA produced in vitro. On the other hand, our results showed that Tdp2 gene knockout did not block CCC DNA formation during HBV infection of permissive human hepatoma cells and did not prevent intracellular amplification of duck hepatitis B virus CCC DNA. These results indicate that although Tdp2 can remove the RT covalently linked to the 5' end of the HBV minus strand DNA in vitro, this protein might not be required for CCC DNA formation in vivo.",
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Cui, X, McAllister, R, Boregowda, R, Sohn, JA, Ledesma, FC, Caldecott, KW, Seeger, C & Hu, J 2015, 'Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair?', PloS one, vol. 10, no. 6, e0128401. https://doi.org/10.1371/journal.pone.0128401

Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair? / Cui, Xiuji; McAllister, Rebecca; Boregowda, Rajeev; Sohn, Ji A.; Ledesma, Felipe Cortes; Caldecott, Keith W.; Seeger, Christoph; Hu, Jianming.

In: PloS one, Vol. 10, No. 6, e0128401, 16.06.2015.

Research output: Contribution to journalArticle

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T1 - Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair?

AU - Cui, Xiuji

AU - McAllister, Rebecca

AU - Boregowda, Rajeev

AU - Sohn, Ji A.

AU - Ledesma, Felipe Cortes

AU - Caldecott, Keith W.

AU - Seeger, Christoph

AU - Hu, Jianming

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Y1 - 2015/6/16

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Cui X, McAllister R, Boregowda R, Sohn JA, Ledesma FC, Caldecott KW et al. Does tyrosyl DNA phosphodiesterase-2 play a role in hepatitis B virus genome repair? PloS one. 2015 Jun 16;10(6). e0128401. https://doi.org/10.1371/journal.pone.0128401