Expression of Autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and Upregulation of the host β-actin gene
Expression of Autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and Upregulation of the host β-actin gene
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DOI：
10.1128/jvi.80.5.2390-2395.2006
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发表时间：
2006-03-01
影响因子：
5.4
通讯作者：
Bando, H
中科院分区：
文献类型：
--
作者：
Fujita, R;Matsuyama, T;Bando, H
研究了苜蓿银纹夜蛾核型多角体病毒（Autographa californica multiple nucleopolyhedrovirus，AcMNPV）在人HeLa 14细胞和仓鼠BHK细胞中的基因表达。DNA微阵列分析和逆转录-PCR检测表明，在接种AcMNPV的HeLa 14细胞和BHK细胞中至少有12个病毒基因转录。用5'端快速扩增cDNA末端的方法检测这些基因在HeLa 14细胞中的转录保真度。ie-1，ie-0和gp 64的转录起始于杆状病毒早期基因基序CAGT，伴随着TATA基序。此外，在Sf 9细胞中观察到的ie-0 mRNA的相同剪接发生在HeLa 14细胞中。虽然pe 38和p6.9的转录起始位点不位于CAGT基序中，但它们中的大多数位于典型的真核RNA聚合酶II启动子结构（常规TATA基序和/或起始子）中。有趣的是，在接种AcMNPV的哺乳动物细胞中，β-肌动蛋白的表达上调。随后使用UV灭活病毒的实验证实了上调，表明该事件不需要病毒产物的从头合成。这些结果表明，AcMNPV基因组作为一个模板，通过通常的感染途径在哺乳动物细胞中的转录，虽然目前还没有证据表明病毒基因的功能表达。
The gene expression of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) was examined in two types of mammalian cells, human HeLa14 and hamster BHK cells. DNA microarray analysis followed by reverse transcription-PCR identified at least 12 viral genes transcribed in both HeLa14 cells and BHK cells inoculated with AcMNPV. 5' rapid amplification of cDNA ends was carried out to examine the transcriptional fidelity of these genes in HeLa14 cells. The transcription of ie-1, ie-0 and gp64 was initiated at a baculovirus early gene motif, CAGT, accompanied by a TATA motif. In addition, the same splicing observed for ie-0 mRNA in Sf9 cells occurred in HeLa14 cells. While the transcription initiation sites for pe38 and p6.9 were not located in the CAGT motif, most of them were in a typical eukaryotic RNA polymerase II promoter structure (a conventional TATA motif and/or an initiator). Interestingly, the expression of beta-actin was upregulated in the mammalian cells inoculated with AcMNPV. Subsequent experiments using UV-inactivated virus confirmed the upregulation, suggesting that de novo synthesis of viral products is not required for the event. These results indicated that the AcMNPV genome acts as a template for transcription in mammalian cells through the usual infection pathway, though there is no evidence for the functional expression of viral genes at present.