Duck Plague Virus pUL48 Protein Activates the Immediate-Early Gene to Initiate the Transcription of the Virus Gene.
Duck Plague Virus pUL48 Protein Activates the Immediate-Early Gene to Initiate the Transcription of the Virus Gene.
复制标题
鸭瘟病毒pUL48蛋白激活即早期基因启动病毒基因转录
DOI：
10.3389/fmicb.2021.795730
复制
发表时间：
2021
影响因子：
5.2
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Zhou T;Fan D;Wang M;Cheng A;Wu Y;Yang Q;Tian B;Jia R;Ou X;Mao S;Sun D;Zhang S;Zhu D;Chen S;Liu M;Zhao XX;Huang J;Gao Q;Yu Y;Zhang L
鸭瘟是由鸭瘟病毒（DPV）引起的一种严重危害水禽养殖业的传染病。α疱疹病毒VP 16蛋白可与立即早期（IE，α）基因启动子上游的特异性顺式作用元件结合，促进IE基因的转录，故又称IE基因的反式诱导因子（α-TIF）。然而，尚未报道关于DPV α-TIF的研究。本研究利用双荧光素酶报告基因检测系统对DPV的pUL 48（HSV-1 VP 16同源物）、转录激活区、靶序列和影响其转录激活的病毒蛋白进行了研究，确定pUL 48为DPV的α-TIF。(1)pUL 48对DPV不同基因启动子的调控表明，pUL 48能激活所有IE基因（ICP 4、ICP 22和ICP 27）的启动子，但不能激活早期和晚期基因的启动子。(2)pUL 48对不同上游长度的ICP 4和ICP 22启动子的活性表明，pUL 48分别作用于ICP 4启动子上游的TAATGA（T）达特元件和ICP 22启动子上游的TAATTATAT元件，从而激活ICP 4和ICP 22启动子。(3)在pUL 48的N端截短不同长度的蛋白质，检测到IE基因的转录激活。结果表明，pUL 48的转录激活区位于N端1-60位氨基酸，其中1-20位氨基酸是其核心区。另外，发现pUL 14、pUL 46和pUL 47显著促进pUL 48的转录激活。进一步研究了pUL 47及其核定位信号的缺失对pUL 48入核和转录激活功能的影响。结果表明，pUL 47可通过其40-50位和768-777位氨基酸的核定位信号，促进pUL 48进入核，从而增强pUL 48的转录激活功能，协同促进病毒基因的转录。
Duck plague caused by the duck plague virus (DPV) is an infectious disease that seriously harms the waterfowl breeding industry. The VP16 protein of α herpesvirus can bind to specific cis-acting elements upstream of the promoter of the immediate-early (IE, α) gene to promote the transcription of the IE gene, so it is also called the trans-inducer of IE gene (α-TIF). However, no studies on DPV α-TIF have been reported. This study investigated the DPV pUL48, a homolog of HSV-1 VP16, transcriptional activation region, target sequence, and viral protein affecting its transcriptional activation using a dual-luciferase reporter gene detection system, and pUL48 was identified as the α-TIF of DPV. (1) The regulation of pUL48 on DPV different gene promoters showed that pUL48 could activate all the promoters of IE genes (ICP4, ICP22, and ICP27) but not the promoters of early and late genes. (2) The activity of pUL48 to ICP4 and ICP22 promoters with different upstream lengths showed that pUL48 activated ICP4 and ICP22 promoters by acting on TAATGA (T) TAT element upstream of ICP4 promoter and TAATTATAT element upstream of ICP22 promoter, respectively. (3) Transcriptional activation of IE gene by truncated proteins of different lengths at the N-terminal of pUL48 was detected. The results showed that the transcriptional activation domain of pUL48 was amino acids 1–60 at the N-terminal, and amino acids 1–20 was its core region. In addition, it was found that pUL14, pUL46, and pUL47 significantly promoted the transcriptional activation of pUL48. The effects of loss of pUL47 and its nuclear localization signal on the nuclear entry and transcriptional activation function of pUL48 were further examined. The results showed that pUL47 could promote the nuclear entry of pUL48 through its nuclear localization signal at positions 40–50 and 768–777 amino acids, thus, enhancing the transcriptional activation function of pUL48 and synergistic promotion of viral gene transcription.