Lu M, Johnson R R, Iatrou K
Department of Medical Biochemistry, University of Calgary, 3330 Hospital Drive, N.W., Calgary, Alberta, T2N 4N1, Canada.
Virology. 1996 Apr 1;218(1):103-13. doi: 10.1006/viro.1996.0170.
Protein IE1 is the product of a baculovirus gene, ie1, that is activated immediately upon entrance of the viral genome into the cell nucleus. This protein was previously shown to be a trans-regulator of viral genes whose products are required for initiation of the infectious cycle including viral DNA replication. To test whether the IE1 protein is also capable of trans-regulating nuclear genes of the host in vitro and in vivo, we transfected the ie1 gene of Bombyx mori nuclear polyhedrosis virus (BmNPV) into silkworm Bm5 tissue culture cells together with expression cassettes directing expression of chloramphenicol acetyl transferase or juvenile hormone esterase under the control of the cytoplasmic actin A3 gene promoter of B. mori. Cotransfection with the ie1 gene resulted in a dramatic increase in the amount of the two enzymes expressed in the transfected cells. The increased enzyme activities correlate with an increased accumulation of the corresponding mRNAs, and the latter is caused by an increase in the rate of transcription directed by the cytoplasmic actin gene promoter. The chromosomal cytoplasmic actin gene of Bm5 cells is also upregulated upon transfection of the cells with the ie1 gene. However, infection of cells with BmNPV does not cause an increase in the level of expression of the endogenous cytoplasmic actin gene. Thus, the effect of IE1 on the transcriptional properties of the cytoplasmic actin gene vary depending on whether IE1 is expressed in isolation or in the context of a viral infection. The trans-activating effects of BmNPV ie1 gene expression on the silkmoth actin promoter are also evident in Spodoptera frugiperda Sf21 and Choristoneura fumiferana Cf1 tissue culture cells. Finally, the ie1 gene of Autographa californica nuclear polyhedrosis virus can substitute for its BmNPV counterpart in all cell lines tested.
蛋白质IE1是杆状病毒基因ie1的产物,该基因在病毒基因组进入细胞核后立即被激活。先前已证明该蛋白是病毒基因的反式调节因子,其产物是感染周期起始所必需的,包括病毒DNA复制。为了测试IE1蛋白在体外和体内是否也能够反式调节宿主的核基因,我们将家蚕核型多角体病毒(BmNPV)的ie1基因与在家蚕细胞质肌动蛋白A3基因启动子控制下指导氯霉素乙酰转移酶或保幼激素酯酶表达的表达盒一起转染到家蚕Bm5组织培养细胞中。与ie1基因共转染导致转染细胞中表达的两种酶的量显著增加。酶活性的增加与相应mRNA积累的增加相关,而后者是由细胞质肌动蛋白基因启动子指导的转录速率增加引起的。用ie1基因转染细胞后,Bm5细胞的染色体细胞质肌动蛋白基因也会上调。然而,用BmNPV感染细胞不会导致内源性细胞质肌动蛋白基因表达水平的增加。因此,IE1对细胞质肌动蛋白基因转录特性的影响取决于IE1是单独表达还是在病毒感染的背景下表达。BmNPV ie1基因表达对家蚕肌动蛋白启动子的反式激活作用在草地贪夜蛾Sf21和云杉卷叶蛾Cf1组织培养细胞中也很明显。最后,苜蓿银纹夜蛾核型多角体病毒的ie1基因可以在所有测试的细胞系中替代其BmNPV对应物。