2009
DOI: 10.1073/pnas.0908365106
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Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function

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 impac… Show more

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Cited by 434 publications
(546 citation statements)
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“…After centrifugation for 1 min at 12,000 rpm at 4˚C, the supernatants were treated with 100 µg/ml DNase I (Promega) for 30 min at 37˚C and then 0.5 mg/ml proteinase K at 50˚C for 2 h. Viral DNA released from lysed cores were extracted with phenol/chloroform, precipitated with ethanol, and dissolved in Tris-EDTA. Nuclear HBV cccDNA was extracted as described (15,21). Simply, transfected cells were lysed in cell lysis buffer [50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.2% NP-40, and 150 mM NaCl].…”
Section: Methodsmentioning
confidence: 99%
“…After centrifugation for 1 min at 12,000 rpm at 4˚C, the supernatants were treated with 100 µg/ml DNase I (Promega) for 30 min at 37˚C and then 0.5 mg/ml proteinase K at 50˚C for 2 h. Viral DNA released from lysed cores were extracted with phenol/chloroform, precipitated with ethanol, and dissolved in Tris-EDTA. Nuclear HBV cccDNA was extracted as described (15,21). Simply, transfected cells were lysed in cell lysis buffer [50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.2% NP-40, and 150 mM NaCl].…”
Section: Methodsmentioning
confidence: 99%
“…HBx carries out many-faceted functions by interacting with a number of cellular proteins in the cytoplasm and nucleus of host cells [10]. Notably, binding of HBx to damage-specific DNA binding protein 1 (DDB1) is essential for HBx to activate HBV transcription [11].…”
Section: Introductionmentioning
confidence: 99%
“…O HBxAg é essencial para a iniciação da transcrição do DNA e manutenção do DNA circular covalentemente fechado (cccDNA) que se acumula no núcleo dos hepatócitos infectados: Este antígeno estimula a acetilação de histonas associadas com cccDNA e é necessário para a transcrição no contexto de infecções por HBV (92). HBxAg promove a expressão dos genes virais recrutando a histona acetilases CBP / p300 e PCAF / GCN5 ao cccDNA (93). Adicionalmente, um estudo recente mostrou que HBx promove a …”
Section: Antígeno Hbx (Hbxag)unclassified