Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function

Laura Belloni, Teresa Pollicino, Francesca De Nicola, Francesca Guerrieri, Giuseppina Raffa, Maurizio Fanciulli, Giovanni Raimondo, Massimo Levrero

Research output: Contribution to journalArticlepeer-review

Abstract

HBV cccDNA, the template for transcription of all viral mRNAs, accumulates in the nucleus of infected cells as a stable episome organized into minichromosomes by histones and non-histone viral and cellular proteins. Using a cccDNA-specific chromatin immunoprecipitation (ChIP)-based quantitative assay, we have previously shown that transcription of the HBV minichromosome is regulated by epigenetic changes of cccDNA-bound histones and that modulation of the acetylation status of cccDNA-bound H3/H4 histones impacts on HBV replication. We now show that the cellular histone acetyltransferases CBP, p300, and PCAF/GCN5, and the histone deacetylases HDAC1 and hSirt1 are all recruited in vivo onto the cccDNA. We also found that the HBx regulatory protein produced in HBV replicating cells is recruited onto the cccDNA minichromosome, and the kinetics of HBx recruitment on the cccDNA parallels the HBV replication. As expected, an HBV mutant that does not express HBx is impaired in its replication, and exogenously expressed HBx transcomplements the replication defects. p300 recruitment is severely impaired, and cccDNA-bound histones are rapidly hypoacetylated in cells replicating the HBx mutant, whereas the recruitment of the histone deacetylases hSirt1 and HDAC1 is increased and occurs at earlier times. Finally, HBx mutant cccDNA transcribes significantly less pgRNA. Altogether our results further support the existence of a complex network of epigenetic events that influence cccDNA function and HBV replication and identify an epigenetic mechanism (i.e., to prevent cccDNA deacetylation) by which HBx controls HBV replication.

Original languageEnglish
Pages (from-to)19975-19979
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number47
DOIs
Publication statusPublished - Nov 24 2009

Keywords

  • HATs
  • HDACs
  • Histone acetylation

ASJC Scopus subject areas

  • General

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