IE1 of Autographa californica Multiple Nucleopolyhedrovirus Activates Low Levels of Late Gene Expression in the Absence of Virus RNA Polymerase.
IE1 of Autographa californica Multiple Nucleopolyhedrovirus Activates Low Levels of Late Gene Expression in the Absence of Virus RNA Polymerase.
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苜蓿银纹夜蛾多核多角体病毒的 IE1 在病毒 RNA 聚合酶缺失的情况下激活低水平的晚期基因表达
DOI：
10.1128/spectrum.03432-22
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发表时间：
2023-02-14
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
已知杆状病毒的早期和晚期基因表达分别依赖于宿主RNA聚合酶II和病毒编码的RNA聚合酶。在这项研究中，我们发现，苜蓿银纹夜蛾多核型多角体病毒（AcMNPV）的重组杆状病毒与个别RNA聚合酶亚基基因删除可以支持低水平的报告基因的表达下的一个典型的晚期基因，vp 39，在转染的Sf 9细胞的启动子的控制。通过病毒基因组文库的多步亚克隆和瞬时表达测定分析，ie 1被鉴定为在不存在病毒RNA聚合酶的情况下负责激活晚期基因表达的唯一病毒基因。此外，IE 1被发现能够激活报告基因表达的启动子额外的晚期基因polh，p6.9，odv-e18，odv-e25，和gp 41，独立于任何额外的病毒因子。病毒基因组中ie 1的缺失消除了晚期基因的表达。IE 1激活的晚期基因表达增强的病毒hr 4 b。它被证明对α-鹅膏蕈碱的抑制不敏感，并且似乎没有稳定的转录起始位点。有人提出，IE 1可能有助于招募新合成的病毒RNA聚合酶的病毒DNA激活低水平的后期基因的预转录，以创建一个适当的DNA构象。重要性已知杆状病毒的晚期基因表达依赖于病毒编码的RNA聚合酶，该聚合酶由四个亚基组成。发现立即早期基因ie 1是病毒早期基因表达、晚期基因表达和DNA复制所必需的。它如何在后期基因表达中发挥作用尚不清楚。在这项研究中，我们发现，AcMNPV IE 1可以激活低水平的基因表达的晚期基因启动子独立于任何其他病毒因素，与非特异性转录起始位点。这一新发现将有助于阐明IE 1在晚期基因表达调控中的作用，并有助于理解晚期基因转录起始的机制。
Early and late gene expressions of baculoviruses have been known to rely on host RNA polymerase II and a virus-encoded RNA polymerase, separately. In this study, we found that Autographa californica multiple nucleopolyhedrovirus (AcMNPV) recombinant bacmids with the individual RNA polymerase subunit genes deleted could support low levels of expression of a reporter gene under the control of the promoter of a typical late gene, vp39, in transfected Sf9 cells. Through multistep subcloning of a genomic library of the virus and transient expression assay analysis, ie1 was identified to be the only viral gene that was responsible for activation of late gene expression in the absence of the viral RNA polymerase. Furthermore, IE1 was found to be capable of activating reporter gene expression from the promoters of additional late genes polh, p6.9, odv-e18, odv-e25, and gp41, independent of any additional viral factors. Deletion of ie1 from the virus genome eliminated late gene expression. The IE1-activated late gene expression was enhanced by the viral hr4b. It was shown to be insensitive to inhibition of α-amanitin and did not appear to have stable transcription start sites. It is proposed that IE1 may serve to recruit newly synthesized viral RNA polymerase to viral DNA by activating low levels of pretranscription of the late genes to create an appropriate DNA conformation. IMPORTANCE The late gene expression of baculovirus has been known to depend on the virus-encoded RNA polymerase, which consists of four subunits. The immediate-early gene ie1 was found to be required for viral early gene expression, late gene expression, and DNA replication. How it functions in late gene expression remains unclear. In this study, we found that AcMNPV IE1 could activate low levels of gene expression from late gene promoters independently of any additional viral factors, with nonspecific transcription start sites. This new finding will shed light on the role of IE1 in the regulation of late gene expression and the understanding of the mechanism of late gene transcription initiation.
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影响因子：
5.4
作者：
Chen, Yun-Ru;Zhong, Silin;Blissard, Gary W.
通讯作者：
Blissard, Gary W.
影响因子：
3.8
作者：
Dai, XJ;Willis, LG;Theilmann, DA
通讯作者：
Theilmann, DA
影响因子：
3.7
作者：
Forsythe, IJ;Shippam, CE;Theilmann, DA
通讯作者：
Theilmann, DA
影响因子：
5.4
作者：
GRULA, MA;BULLER, PL;WEAVER, RF
通讯作者：
WEAVER, RF
DOI：
10.1073/pnas.91.23.11212
发表时间：
1994-11-08
影响因子：
11.1
作者：
KOOL, M;AHRENS, CH;VLAK, JM
通讯作者：
VLAK, JM