[Frontiers in Bioscience, Landmark, 25, 710-721, Jan 1, 2020]

ADAR1 promotes HBV replication through its deaminase domain

Lin Yuan1,2, Qianying Jia3, Shenyong Yang1, Nur Fazleen Binti Idris1, Yimin Li1, Yang Wang1, Ya Zhao2, Jieli Hu4, Yuan Hu4, Ailong Huang4, Zeng Tu1, 4

1Department of Microbiology, College of Basic Medical Sciences/Molecular Medicine and Tumor Research Center, Chongqing Medical University, Chongqing 400016, China, 2Department of Clinical Laboratory, The First Hospital of Xi'an, Xi'an 710002, China, 3Department of Infectious Diseases, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China, 4Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
    3.1. Cell lines
    3.2. Plasmids, small interfering RNAs, and transfection
    3.3. Western blot analysis
    3.4. Quantitative PCR analysis
    3.5. Southern blotting and qPCR detection for HBV total DNA
    3.6. Statistical analysis
4. Results
    4.1. ADAR1 knockdown down-regulated HBV replication
    4.2. Over-expressed ADAR1 enhanced HBV replication
    4.3. ADAR1 increased the replication of HBV independent of genotypes
    4.4. ADAR1 facilitated HBV replication through its editing function
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

Adenosine deaminase acting on RNA1 (ADAR1) is an RNA editing enzyme that modulates the replication of several viruses. Here, we provide evidence showing that ADAR1 stimulates hepatitis B virus (HBV) DNA replication in hepatocellular carcinoma cell lines that are transiently or stably-transfected with HBV. We show that overexpression of ADAR1 promotes the replication of all four HBV genotypes (A, B, C, and D). Furthermore, analysis by mutagenesis shows that RNA editing region, and to a lesser content, RNA binding region of ADAR is responsible for the promotion of replication. Together, these data show that ADAR1 stimulates HBV replication.

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Key Words: Adenosine Deaminase, RNA1, ADAR1, Hepatitis B virus, RNA Editing, Replication

Send correspondence to: Ailong Huang or Zeng TU, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, NO.1 YiXueYuan Road, Yuzhong District, Chongqing, 400016, P.R. China, Tel: 862368486780, Fax: 862368486780, E-mail: tuzeng@cqmu.edu.cn