Altering of host larval (Spodoptera exigua) calcineurin activity in response to ascovirus infection
Altering of host larval (Spodoptera exigua) calcineurin activity in response to ascovirus infection
复制标题
改变宿主幼虫（甜菜夜蛾）钙调神经磷酸酶活性以响应子囊病毒感染
DOI：
10.1002/ps.5615
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发表时间：
2019
影响因子：
4.1
通讯作者：
Guo-Hua Huang
中科院分区：
文献类型：
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作者：
Huan Yu;Lei He;Zi-Qi Li;Ni Li;Yi-Yi Ou-Yang;Guo-Hua Huang
背景钙调神经磷酸酶(CaN)参与多种细胞过程和钙依赖的信号转导途径。根据我们以前的转录组研究，甜菜夜蛾幼虫在感染HvAV-3h后，数千个转录本下调，而甜菜夜蛾钙调神经磷酸酶基因(SeCaN)显著上调。为了了解甜菜夜蛾幼虫在HvAV-3h感染过程中SeCaN的调控，对CaN A亚基(SeCaN-Suba)和CaN结合蛋白(SeCaN-BP)的功能进行了分析。结果体外实验表明，细菌表达的SeCaN-SuBA是一种酸性磷酸酶，而纯化的SeCaN-BP没有检测到磷酸酶活性。HvAV-3h感染甜菜夜蛾幼虫后，SeCaN-SuBA转录水平上调，CaN活性明显增强。有趣的是，只有在感染HvAV-3h的幼虫中才能检测到SeCaN-BP转录本。进一步的免疫印迹结果与qPCR的结果一致，表明HvAV-3h感染导致SeCaN-Suba表达上调和SeCaN-BP的出现。免疫共沉淀法检测到裂解的SeCaN-Suba与SeCaN-BP之间存在相互作用，SeCaN-BP在果蝇Sf9细胞中的表达有助于提高SeCaN-Suba的活性。对CaN抑制剂的进一步研究表明，HvAV-3h。对CaN抑制剂的进一步研究表明，对寄主幼虫CaN活性的抑制也可以抑制HvAV-3h的病毒复制。结论HvAV-3h感染后细胞内CaN活性升高可能与SeCaN-Suba表达上调和SeCaN-BP表达上调有关，而CaN活性升高对囊泡病毒复制起重要作用。(三)2019年化学工业学会
BACKGROUND Calcineurin (CaN) is involved in numerous cellular processes and Ca2+-dependent signal transduction pathways. According to our previous transcriptome studies, thousands of host larval (Spodoptera exigua) transcripts were downregulated after the infection of Heliothis virescent ascovirus 3h (HvAV-3h), while the Spodoptera exigua calcineurin genes (SeCaNs) were significantly upregulated. To understand the regulation of SeCaNs in S. exigua larvae during the infection of HvAV-3h, the functions of CaN subunit A (SeCaN-SubA) and CaN binding protein (SeCaN-BP) were analysed. RESULTS The in vitro assays indicated that the bacterial expressed SeCaN-SubA is an acid phosphatase, but no phosphatase activity was detected with the purified SeCaN-BP. The transcription level of SeCaN-SubA was upregulated after HvAV-3h infection and the CaN activity was significantly increased after HvAV-3h infection in S. exigua larvae. Interestingly, the SeCaN-BP transcripts were only detectable in the HvAV-3h infected larvae. Further immunoblotting results consistently agree with those obtained by qPCR, indicating that the infection of HvAV-3h causes the upregulated expression of SeCaN-SubA and the appearance of SeCaN-BP. An interaction between the cleaved SeCaN-SubA and SeCaN-BP was detected by co-immunoprecipitation assays, and the expression of SeCaN-BP in Spodoptera frugiperda-9 (Sf9) cells can help to increase the CaN activity of SeCaN-SubA. Further investigations with CaN inhibitors suggested that HvAV-3h. Further investigations with CaN inhibitors suggested that the inhibition on host larval CaN activity can also inhibit the viral replication of HvAV-3h. CONCLUSION The increase in CaN activity caused by HvAV-3h infection might be due to the upregulation of SeCaN-SubA and the induced expression of SeCaN-BP, and increased CaN activity is essential for ascoviral replication. (c) 2019 Society of Chemical Industry