Rabies virus phosphoprotein P5 binding to BECN1 regulates self-replication by BECN1-mediated autophagy signaling pathway
Rabies virus phosphoprotein P5 binding to BECN1 regulates self-replication by BECN1-mediated autophagy signaling pathway
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DOI：
10.1186/s12964-020-00644-4
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发表时间：
2020-09-18
影响因子：
8.4
通讯作者：
Zhou, Jiyong
中科院分区：
文献类型：
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作者：
Liu, Juan;Liao, Min;Zhou, Jiyong
背景：据报道，狂犬病毒（RABV）编码5种磷酸化蛋白(P)，参与病毒基因组复制、轴突运输、氧化应激、干扰素拮抗和自噬诱导。然而，人们对不同P蛋白的功能知之甚少。方法：采用免疫荧光染色和western blot检测自噬活性、环状结构的形成，采用共定位法检测BECN1与P的相互作用。采用QRT-PCR和TCID50法检测RABV的复制水平。采用小干扰RNA检测自噬信号通路。结果：我们发现P5通过P5的173-222个氨基酸残基附着在BECN1环状结构的n端残基1-139 （beclin1）上。随后，我们发现p5诱导的自噬体不与溶酶体融合。becn1沉默不能恢复P5过表达诱导的RABV复制促进。在机制上，RABV蛋白P δ N82 （P5）通过becn1介导的信号通路诱导不完全自噬。结论：我们的数据表明P5与BECN1环的结合通过诱导BECN1信号通路依赖的不完全自噬促进RABV的复制，这为RABV抗病毒药物提供了潜在的靶点。
Background: Rabies virus (RABV) is reported to encode five phosphoproteins (P), which are involved in viral genomic replication, axonal transport, oxidative stress, interferon antagonism, and autophagy induction. However, the functions of the different P proteins are poorly understood.Methods: Immunofluorescence staining and western blot were performed to detect the autophagy activity, the form of ring-like structure, and the colocalization of BECN1 and P. Co-immunoprecipitation was performed to detect the interaction between P and BECN1. QRT-PCR and TCID50 assay were performed to detect the replication level of RABV. Small interfering RNA was used to detect the autophagy signaling pathway.Results: We found that P5 attaches to N-terminal residues 1-139 of BECN1 (beclin1) on the BECN1 ring-like structure through amino acid residues 173-222 of P5. Subsequently, we found that P5-induced autophagosomes did not fuse with lysosomes.Becn1silencing did not recover P5 overexpression-induced promotion of RABV replication. Mechanistically, RABV protein P Delta N82 (P5) induced incomplete autophagy via the BECN1-mediated signaling pathway.Conclusions: Our data indicate that P5 binding to the BECN1 ring benefits RABV replication by inducing BECN1 signaling pathway-dependent incomplete autophagy, which provides a potential target for antiviral drugs against RABV.