Yan Liu

Publications (3)9.27 Total impact

• Article: Human microRNA hsa-miR-296-5p suppresses Enterovirus 71 replication by targeting the viral genome.

  Zhenhua Zheng, Xianliang Ke, Meng Wang, Siyi He, Qian Li, Caishang Zheng, Zhenfeng Zhang, Yan Liu, Hanzhong Wang
  [show abstract] [hide abstract]
  ABSTRACT: Enterovirus 71 (EV71) has emerged as a major cause of neurological disease following the near eradication of poliovirus. Accumulating evidence suggest that mammalian microRNAs (miRNAs), a class of noncoding RNAs of 18-23 nucleotides (nt) with important regulatory roles in many cellular processes, participate in host anti-viral defenses. However, roles of miRNAs in EV71 infection and pathogenesis are still unclear. Here, hsa-miR-296-5p expression was significantly increased in EV71-infected human cells. As determined by virus titration, quantitative real-time PCR (qRT-PCR) and Western blotting, overexpression of hsa-miR-296-5p inhibited, while inhibition of endogenous hsa-miR-296-5p facilitated, EV71 infection. Additionally, two potential hsa-miR-296-5p targets (nt 2115-2135 and nt 2896-2920) located in the EV71 genome (BrCr strain) were bioinformatically predicted and validated by luciferase reporter assays and Western blotting. Genomic alignment of various EV71 strains revealed synonymous mutations in hsa-miR-296-5p target sequences. Furthermore, introduction of synonymous mutations into the EV71 BrCr genome by site-directed mutagenesis impaired the viral inhibitory effects of hsa-miR-296-5p and facilitated mutant virus infection. Meanwhile, compensatory mutations in corresponding hsa-miR-296-5p target sequences of the EV71 HeN strain (GenBANK accession: JN256064) restored the inhibitory effects of the miRNA. These results indicate that hsa-miR-296-5p inhibits EV71 replication by targeting the viral genome. Our findings support the notion that cellular miRNAs can inhibit virus infection and that the virus mutates to escape suppression by cellular miRNAs.
  Journal of Virology 03/2013; · 5.40 Impact Factor
• Article: Methionine-101 from one strain of H5N1 NS1 protein determines its IFN-antagonizing ability and subcellular distribution pattern.

  Jin Meng, Zhenfeng Zhang, Zhenhua Zheng, Yan Liu, Hanzhong Wang
  [show abstract] [hide abstract]
  ABSTRACT: Influenza A virus NS1 protein has developed two main IFN-antagonizing mechanisms by inhibiting retinoic-acid-inducible gene I (RIG-I) signal transduction, or by suppressing cellular pre-mRNA processing through binding to cleavage and polyadenylation specific factor 30 (CPSF30). However, the precise effects of NS1 on suppressing type I IFN induction have not been well characterized. Here we report that compared with PR/8/34 NS1, which is localized partially in the cytoplasm and has strong IFN-antagonizing ability via specifically inhibiting IFN-β promoter activity, H5N1 NS1 has strikingly different characteristics. It mainly accumulates in the nucleus of transfected cells and exerts rather weak IFN-counteracting ability through suppression of the overall gene expression. The M101I mutation of H5N1 NS1, namely H5-M101I, fully reversed its functions. H5-M101I gained the ability to specifically inhibit IFN-β promoter activity, translocate to the cytoplasm, and release CPSF30. The previously reported NES (nuclear export signal) (residues 138-147) was unable to lead H5N1 NS1 to translocate. This suggests that other residues may serve as a potent NES. Findings indicated that together with leucine-100, methionine-101 enhanced the regional NES. In addition, methionine-101 was the key residue for the NS1-CPSF30 interaction. This study reveals the importance of methionine-101 in the influenza A virus life cycle and may provide valuable information for antiviral strategies.
  Science China. Life sciences 11/2012; · 2.02 Impact Factor
• Article: Helicoverpa armaigera nucleopolyhedrovirus ORF50 is an early gene not essential for virus propagation in vitro and in vivo.

  Yao Chen, Fangliang Zheng, Ling Tao, Zhenhua Zheng, Yan Liu, Hanzhong Wang
  [show abstract] [hide abstract]
  ABSTRACT: Homologs of Helicoverpa armigera nucleopolyhedrovirus ORF50 (HA50) are found in most alphabaculoviruses, but their functions remain unknown. Here, we characterized whether Ha50 is indispensable for virus progration. Ha50 transcript was first detected at 3 h post-infection from HearNPV-infected HzAM1 cells. 3'RACE analysis showed that Ha50 transcript was polyadenlylated. 5'RACE analysis revealed two transcription initiation sites, one of which was mapped to the canonical baculovirus early transcription initiator motif CAGT. HA50 protein could be detected from infected cells harvested at 12 h post-infection. Transient expression assays showed that GFP-fused HA50 localized in the cytoplasm and nucleus of HzAM1 cells with or without superinfection. To further examine the role of Ha50 in the virus life cycle, a Ha50 knockout bacmid and a repair bacmid carrying Ha50 under the control of its native promoter elements were constructed using bacmid technology. One-step growth curve analysis showed that the kinetics of infectious budded virus production of Ha50 knockout virus was similar to that of the parental virus or the repair virus. Analysis of the expression of viral early protein IE-1, late protein VP39 and very late protein suggested that viral protein expression was not affected by Ha50 inactivation. Electron microscopy revealed that HaBacΔ50-PH-G occluded viruses (ODVs) and occlusion bodies were indistinguishable from those of the wild-type virus. Similarly, bioassays showed no significant difference in the LC(50) values between Ha50 deletion virus and wild-type virus. Our results together demonstrate that Ha50 is an early gene dispensable for virus propagation in vitro and in vivo.
  Virus Genes 05/2012; 45(1):149-60. · 1.85 Impact Factor