Braunagel S C, Elton D M, Ma H, Summers M D
Texas Agricultural Experiment Station, Department of Entomology, Texas A&M University, College Station 77843, USA.
Virology. 1996 Mar 1;217(1):97-110. doi: 10.1006/viro.1996.0097.
An Autographa californica nuclear polyhedrosis virus gene encoding an occlusion-derived virus (ODV) envelope protein of 56 kDa was identified and sequenced. Transcription initiates from a conserved baculovirus late motif (ATAAG) with transcripts detected from 16 through 72 hr p.i. The protein is detected in infected cell extracts from 36 hr p.i. Western blot assay of ODV, BV, viral envelope, and nucleocapsid preparations coupled with immunoelectron microscopy reveal that this protein localizes to the ODV envelope. This protein is named ODV-E56 to identify its viral origin, envelope location, and apparent molecular weight. ODV-E56 is enriched in viral induced intranuclear microvesicles as determined by immunogold labeling. A mutant was constructed with the C-terminal portion of the protein replaced with beta-galactosidase. The fusion protein, E56-beta-gal, locates to the viral nucleocapsids and not to the ODV envelope or intranuclear microvesicles. This suggests that the signals necessary for transport and/or retention into these structures lies within the C-terminal portion of ODV-E56. Additionally, both ODV-E56 and E56-beta-gal are enriched in electron dense regions that cluster around the inner nuclear membrane and within the nucleoplasm.
鉴定并测序了苜蓿银纹夜蛾核型多角体病毒中一个编码56 kDa的包涵体衍生病毒(ODV)包膜蛋白的基因。转录起始于一个保守的杆状病毒晚期基序(ATAAG),在感染后16至72小时可检测到转录本。在感染后36小时的感染细胞提取物中可检测到该蛋白。对ODV、BV、病毒包膜和核衣壳制剂进行蛋白质印迹分析,并结合免疫电子显微镜观察,结果表明该蛋白定位于ODV包膜。为了确定其病毒来源、包膜位置和表观分子量,将该蛋白命名为ODV-E56。通过免疫金标记确定,ODV-E56在病毒诱导的核内微泡中富集。构建了一个突变体,将该蛋白的C末端部分替换为β-半乳糖苷酶。融合蛋白E56-β-gal定位于病毒核衣壳,而不是ODV包膜或核内微泡。这表明运输和/或保留在这些结构中所需的信号位于ODV-E56的C末端部分。此外,ODV-E56和E56-β-gal都富集在围绕内核膜和核质内聚集的电子致密区域。