Expression and localization of LEF-11 in Autographa californica nucleopolyhedrovirus-infected Sf9 cells.
Expression and localization of LEF-11 in Autographa californica nucleopolyhedrovirus-infected Sf9 cells.
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DOI：
10.1099/0022-1317-82-9-2289
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发表时间：
2001-09
期刊：
影响因子：
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通讯作者：
Guangyun Lin;J. Slack;G. Blissard
中科院分区：
文献类型：
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作者：
Guangyun Lin;J. Slack;G. Blissard
苜蓿银纹夜蛾核型多角体病毒（Autographa californica multicapsid nucleopolyhedrovirus，AcMNPV）lef-11基因是支持AcMNPV晚期启动子瞬时表达最佳水平所必需的。lef-11基因是不寻常的，因为它重叠上游（orf 38）和下游（pp 31）基因。在这项研究中，LEF-11的表达和细胞定位进行了检查。lef-11转录本在感染后4 - 36 h检测到。lef-11 mRNA起始于lef-11翻译起始密码子上游196 nt处，位于orf 38基因上游。该相对较长的5'上游区编码一个潜在的小的上游开放阅读框（ORF），其为58个氨基酸，与lef-11 ORF重叠。lef-11 mRNA的3'末端与下游pp 31基因的mRNA共末端。使用亲和纯化的抗LEF-11抗体，发现LEF-11表达水平在感染后约8至24小时之间最大，尽管LEF-11可以迟至感染后72小时检测到。使用免疫荧光显微镜，确定LEF-11定位于感染细胞核的致密区域，这与其作为可能的晚期转录因子的作用一致。
The Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) lef-11 gene was found previously to be necessary to support optimal levels of transient expression from an AcMNPV late promoter. The lef-11 gene is unusual in that it overlaps both upstream (orf38) and downstream (pp31) genes. In this study, the expression and cellular localization of LEF-11 were examined. The lef-11 transcripts were detected from 4 to 36 h post-infection (p.i.). The 1.5 kb lef-11 mRNA initiates 196 nt upstream of the lef-11 translation initiation codon, within the upstream orf38 gene. This relatively long 5' upstream region encodes a potential small upstream open reading frame (ORF) of 58 amino acids that overlaps the lef-11 ORF. The 3' end of the lef-11 mRNA was mapped as co-terminal with mRNAs from the downstream pp31 gene. Using affinity purified anti-LEF-11 antibodies, levels of LEF-11 expression were found to be maximal between approximately 8 and 24 h p.i., although LEF-11 could be detected as late as 72 h p.i. Using immunofluorescence microscopy, it was determined that LEF-11 localized to dense regions of infected cell nuclei, consistent with its role as a possible late transcription factor.