The Inhibition of IFN Induce SAMHD1 to the Replication of HBV in Huh7.0 Cells

doi:10.13523/j.cb.20190301

China Biotechnology

2019, Vol. 39

Issue (3): 1-6 DOI: 10.13523/j.cb.20190301

The Inhibition of IFN Induce SAMHD1 to the Replication of HBV in Huh7.0 Cells

Miao QIAO,Jie HU,Bin-li MAO,Si-die PI,Yuan HU

Key Laboratory of Molecular Biology on Infectious Disease, Ministry of Education,Chongqing Medical University, Chongqing 400016, China

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Abstract

Objective: To analyze the mechanism of the inhibition of IFNs induce SAMHD1 to hepatitis B virus(HBV)in liver cancer cells Huh7.0. Methods: (1) Recombiant expression plasmids of SAMHD1 (sterile alpha motif and histidine/ aspartic acid domain-containing protein 1) mutants that were defective in dNTPase (deoxynucleoside triphosphate triphosphohydrolase) activity and its nuclear location mutation were constructed. (2) The mRNA and protein levels of SAMHD1 in Huh7.0 cells by IFNs treatment were measured by real time and Western blot. (3) HBV core-associated DNA levels in transfected Huh7.0 cells were measured by Southern blot. (4) The location of SAMHD1 and its nuclear mutation in the Huh7.0 cells was measured by immunofluorescence. Results: (1) Both the RNA and protein levels of SAMHD1 were upregulated by interferons. (2) The antiviral function of SAMHD1 was lost when knocking down the expression of SAMHD1 by siRNA. (3) SAMHD1 lost its restriction towards HBV after mutating its NLS signal. Conclusion: IFNs can upregulate the expressing of SAMHD1 in Huh7 cells. SAMHD1 is mainly located in the nucleus and its restriction towards HBV dependent on its location.

Key words： Hepatitis B virus Interferons SAMHD1 Inhibition

Received: 12 September 2018 Published: 12 April 2019

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Cite this article:

Miao QIAO,Jie HU,Bin-li MAO,Si-die PI,Yuan HU. The Inhibition of IFN Induce SAMHD1 to the Replication of HBV in Huh7.0 Cells. China Biotechnology, 2019, 39(3): 1-6.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20190301 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I3/1

Table 1 Sequence of primers for site-directed mutagenesis and siRNA target for SAMHD1

Fig.1 Interferon α、β、γ induce SAMHD1 expression in Huh7.0 cells

Fig.2 Effect of SAMHD1 knockdown on inhibition of IFN α、β toward HBV RC:Relaxed circular DNA;DL:Double-stranded linear DNA;SS:Single-stranded DNA

Fig.3 Distribution of SAMHD1 wide type and NLS deletion mutant

Fig.4 Inhibition of SAMHD1 and its NLS mutation on HBV RC DNA:Relaxed circular DNA;DL DNA:Double-stranded linear DNA;SS DNA:Single-stranded DNA


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