The 2A2 protein of Duck hepatitis A virus type 1 induces apoptosis in primary cell culture
The 2A2 protein of Duck hepatitis A virus type 1 induces apoptosis in primary cell culture
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鸭甲型肝炎病毒1型2A2蛋白诱导原代细胞培养细胞凋亡
DOI：
10.1007/s11262-016-1364-4
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发表时间：
2016-12-01
期刊：
影响因子：
1.6
通讯作者：
Chen, Xiaoyue
中科院分区：
文献类型：
--
作者：
Cao, Jingyu;Ou, Xumin;Chen, Xiaoyue
鸭甲型肝炎病毒1型（DHAV-1） 2A2pro是DHAV血清型中最高度保守的病毒蛋白之一。然而，其对宿主细胞的影响尚不清楚。基于多序列比对和同源建模分析，我们预测DHAV-1 2A2pro为gtpase样蛋白。转染表达DHAV-1 2A2的重组质粒后，细胞显示细胞核碎片化、染色质凝聚、寡核小体大小的DNA阶梯和阳性末端脱氧核苷酸转移酶介导的dutp -生物素缺口端标记染色；因此，细胞死亡具有凋亡的特征。用荧光素Annexin V-FITC和PI对细胞进行染色，流式细胞术和荧光显微镜可以对非凋亡细胞、早期凋亡细胞、晚期凋亡/坏死细胞和死亡细胞进行区分和定量分析。凋亡细胞比例逐渐增加，转染48 h后达到最大值。总之，这种gtpase样蛋白诱导的细胞凋亡可能参与了DHAV-1的发病机制。
Duck hepatitis A virus type 1, (DHAV-1) 2A2pro, is one of the most highly conserved viral proteins within the DHAV serotypes. However, its effect on host cells is unclear. We predicted that DHAV-1 2A2pro was a GTPase-like protein based on the results of multiple sequence alignment and homologous modeling analysis. Upon transfection of a recombinant plasmid expressing DHAV-1 2A2, cells displayed fragmented nuclei, chromatin condensation, oligonucleosome-sized DNA ladder, and positive terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining; hence, cell death has the characteristics of apoptosis. By staining cells with fluorescein Annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze nonapoptotic cells, early apoptotic cells, late apoptotic/necrotic cells, and dead cells through flow cytometry and fluorescence microscopy. The percentage of apoptotic cells gradually increased and reached a maximum after 48 h of transfection. In conclusion, apoptosis induced by this GTPase-like protein may contribute to DHAV-1 pathogenesis.