Pular para o conteúdo
Merck

DHX9 interacts with APOBEC3B and attenuates the anti-HBV effect of APOBEC3B.

Emerging microbes & infections (2020-02-15)
Yanmeng Chen, Bocun Shen, Xiaochuan Zheng, Quanxin Long, Jie Xia, Yao Huang, Xuefei Cai, Deqiang Wang, Juan Chen, Ni Tang, Ailong Huang, Yuan Hu
RESUMO

Hepatitis B virus (HBV) is a partially double-stranded DNA virus that replicates by reverse transcription. We previously demonstrated that the host restriction factor-APOBEC3B (A3B) inhibited HBV replication which was dependent on its deaminase activity during reverse transcription. However, the host factors involved in the process of regulating the anti-HBV function of A3B are less known. In this research, to obtain a comprehensive understanding of the interaction networks of A3B, we conducted coimmunoprecipitation and mass spectrometry to identify A3B-interacting proteins in the presence of HBV. By this approach, we determined that DExD/H-box helicase 9 (DHX9) suppressed the anti-HBV effect of A3B, and this suppression was dependent on their interaction. Although DHX9 did not affect the deamination activity of A3B in vitro assay or the viral DNA editing of A3B in HepG2-NTCP cells that support HBV infection, it inhibited the binding of A3B with pgRNA. These data suggest that DHX9 can interact with A3B and attenuate the anti-HBV efficacy of A3B.Abbreviations: 3D-PCR: differential DNA denaturation PCR; APOBEC3: apolipoprotein B mRNA-editing catalytic polypeptide 3; cccDNA: covalently closed circular DNA; co-IP: coimmunoprecipitation; DDX: DExD-box RNA helicases; HBc: HBV core protein; HBV: hepatitis B virus; HepAD38: HepG2 cell line stably transfected with HBV DNA; HepG2-NTCP: HepG2 cell line stably transfected with Na+/taurocholate cotransporter polypeptide; Huh7: human hepatoma cell line; pgRNA: pregenomic RNA; PPI: protein-protein interactions; RC DNA: relaxed circular DNA.

MATERIAIS
Número do produto
Marca
Descrição do produto

Sigma-Aldrich
Magna RIP® RNA-Binding Protein Immunoprecipitation Kit, RNA Immunoprecipitation (RIP) Kit containing all necessary reagents to perform 12 individual RNA-binding protein immunoprecipitation (RIP) reactions using protein A/G magnetic beads.
Roche
DIG-High Prime DNA Labeling and Detection Starter Kit II, sufficient for 12 labeling reactions (10 ng to 3 μg per assay), sufficient for 24 blots (blots of 100 cm2)
Millipore
Agarose de proteína G, vazão rápida, Protein G Agarose, Fast Flow suitable for medium and low pressure chromatography of IgG from mouse, sheep, and rabbit, and for immunoprecipitations.