Bilimoria S L, Demirbag Z, Ng H
Department of Biological Sciences, Texas Tech University, Lubbock 79409-3131.
SAAS Bull Biochem Biotechnol. 1993;6:1-7.
We examined the differential replication of Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV; Family: Baculoviridae) in Spodoptera frugiperda (SF) and Bombyx mori (BM) cell lines. Our previous studies have shown that infection of BM cells is abortive; infectious progeny is not produced, and although virogenic stroma is produced, complete virions or inclusion bodies are not assembled. We designed the present study to determine the extent of infected-cell-specific polypeptides (ICSPs) and viral DNA synthesis in the abortive infection and to see if restriction of virus replication in BM cells is due to a block at the transcriptional level. Pulse-labeling studies showed that a majority of the ICSPs detected in productive SF cells were not synthesized in BM cells. Many of these ICSPs were virion components. Viral DNA replicated in BM cells but more slowly and at a much lower level than in SF cells. Northern blot analysis showed that i) AcMNPV immediate-early gene (IE-1) was transcribed in BM cells at higher levels than in SF cells; ii) a delayed-early gene (dnapol) and a late gene (cap) were transcribed at much lower levels in the abortive infection; and iii) a very late gene (polh) was transcribed at extremely low levels in BM cells. We conclude that virus replication in BM cells is primarily restricted during or prior to the delayed-early stage and occurs at the transcriptional level. It appears that the late and very late effects observed are consequences of the primary block.
我们研究了苜蓿银纹夜蛾多粒包埋核型多角体病毒(AcMNPV;杆状病毒科)在草地贪夜蛾(SF)和家蚕(BM)细胞系中的差异复制情况。我们之前的研究表明,BM细胞的感染是流产性的;不会产生有感染性的子代病毒,并且尽管会产生病毒发生基质,但完整的病毒粒子或包涵体并未组装形成。我们开展本研究以确定流产性感染中感染细胞特异性多肽(ICSPs)和病毒DNA合成的程度,并探究BM细胞中病毒复制受限是否是由于转录水平的阻滞。脉冲标记研究表明,在产生子代病毒的SF细胞中检测到的大多数ICSPs在BM细胞中并未合成。这些ICSPs中的许多都是病毒粒子成分。病毒DNA在BM细胞中能够复制,但比在SF细胞中更慢且水平更低。Northern印迹分析表明:i)AcMNPV立即早期基因(IE-1)在BM细胞中的转录水平高于SF细胞;ii)在流产性感染中,一个延迟早期基因(dnapol)和一个晚期基因(cap)的转录水平要低得多;iii)一个极晚期基因(polh)在BM细胞中的转录水平极低。我们得出结论,BM细胞中的病毒复制主要在延迟早期阶段或之前受到限制,并且发生在转录水平。似乎观察到的晚期和极晚期效应是主要阻滞的结果。