Functional characterization of Bombyx mori nucleopolyhedrovirus late gene transcription and genome replication factors in the non-permissive insect cell line SF-21.
Functional characterization of Bombyx mori nucleopolyhedrovirus late gene transcription and genome replication factors in the non-permissive insect cell line SF-21.
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家蚕核多角体病毒晚期基因转录和基因组复制因子在非许可昆虫细胞系 SF-21 中的功能表征。
DOI：
10.1016/j.virol.2005.12.017
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发表时间：
2006
期刊：
影响因子：
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通讯作者：
Passarelli,ALorena
中科院分区：
文献类型：
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作者：
Berretta,MarceloF;Deshpande,Mandar;Crouch,ErinA;Passarelli,ALorena
比较了苜蓿银纹夜蛾核型多角体病毒（Autographa californica Nucleopolyhedrovirus，AcMNPV）和家蚕核型多角体病毒（BmNPV）在昆虫源细胞系SF-21中晚期基因转录和DNA复制机制的能力。已经确定的是，19个AcMNPV晚期表达因子（lefs）在SF-21细胞中刺激实质水平的晚期基因启动子活性。因此，我们构建了一组克隆含有BmNPV同源物的AcMNPV lefs的组成型果蝇热休克70蛋白启动子的控制下，并测试了它们的能力，激活SF-21细胞中的AcMNPV晚期启动子报告基因盒。我们测试了单个或预测的BmNPV lefs功能组在瞬时晚期基因表达测定中成功取代相应AcMNPV基因的潜力。我们发现，大多数，但不是所有的，BmNPV lefs能够完全或部分取代相应的AcMNPV同源物的上下文中的其余AcMNPV lefs的BmNPV p143，即-2，p35的例外。BmNPV p143不能支持晚期基因表达或在AcMNPV或BmNPV LEF-3（一种P143核穿梭因子）存在下被导入细胞核。我们的研究结果表明，宿主特异性因素可能会影响同源蛋白的功能。
We compared the abilities of late gene transcription and DNA replication machineries of the baculoviruses Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) in SF-21 cells, an insect-derived cell line permissive for AcMNPV infection. It has been well established that 19 AcMNPV late expression factors (lefs) stimulate substantial levels of late gene promoter activity in SF-21 cells. Thus, we constructed a set of clones containing the BmNPV homologs of the AcMNPV lefs under control of the constitutive Drosophila heat shock 70 protein promoter and tested their ability to activate an AcMNPV late promoter–reporter gene cassette in SF-21 cells. We tested the potential of individual or predicted functional groups of BmNPV lefs to successfully replace the corresponding AcMNPV gene(s) in transient late gene expression assays. We found that most, but not all, BmNPV lefs were able to either fully or partially substitute for the corresponding AcMNPV homolog in the context of the remaining AcMNPV lefs with the exception of BmNPV p143, ie-2, and p35. BmNPV p143 was unable to support late gene expression or be imported into the nucleus of cells in the presence of the AcMNPV or the BmNPV LEF-3, a P143 nuclear shuttling factor. Our results suggest that host-specific factors may affect the function of homologous proteins.