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| APOBEC3A Inhibited HBV Replication Dependent Its Deamination |
| CHEN Yan-meng, HUANG Yao, HU Jie, HU Yuan |
| Key Laboratory of Molecular Biology on Infectious Disease, Ministry of Education, Chongqing Medical University, Chongqing 400016, China |
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Abstract Purpose:To investigate the mechanism of APOBEC3A on HBV replication. Methods:(1) The cytotoxicity effect of exogenous APOBEC3A in HuH7 cells was evaluated by MTT assay. (2) The location of APOBEC3A in the HuH7 cells was measured by immunofluorescence, and the interaction of APOBEC3A with virus core particles was further confirmed by IP. (3) The relative levels of HBV replication parameters:including HBsAg in the supernatant, HBV DNA in virus core particles and intranuclear cccDNA were measured by ELISA and specific real-time PCR in the presence of exogenous APOBEC3A. (4) The deamination of HBV DNA in core particles was analyzed by 3D PCR. Results:(1) The expression of exogenous APOBEC3A had no effect on cell viability of HuH7 cells. (2)APOBEC3A was mainly located in the nucleus, but APOBEC3A combined with virus core particles to deaminate DNA during the process of HBV reverse transcription. (3) After co-transfection of HBV replication plasmid and APOBEC3A expression plasmid, the levels of HBsAg in the supernatant, HBV DNA in particles, and cccDNA were significantly decreased by 80%, 64% and 82%, respectively.(4) Hypermutations of G-A accumulated in the HBV minus strand DNA and C-T in the plus strand DNA of core particles in the presence of exogenous APOBEC3A were confirmed by 3D PCR and sequencing. Conclusion:APOBEC3A can interact with virus core particles, target HBV single strand DNA during the process of HBV reverse transcription, and inhibit HBV replication dependent its deamination.
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Received: 06 August 2015
Published: 22 December 2015
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