Role of AcMNPV IE0 in baculovirus very late gene activation

IE0 is the only known baculovirus protein that is produced by splicing. In this study, we have explored the role of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) IE0 and its interaction with IE1 in the activation of very late gene expression from the polyhedrin promoter using transient assays. IE0 is co-expressed with IE1 throughout infection up to late times post-infection (p.i.) but shows peak levels of expression at early times. Significant changes in the ratios of the relative levels of IE0 to IE1 were observed throughout the course of infection. To study IE0 in the absence of IE1, we constructed a plasmid pAc-IE0(M-->A) that expressed only IE0. This was due to a mutation of the internal AUG that prevented translation of IE1 from the ie0 mRNA. Both IE0 and IE0(M-->A) were able to replace IE1 in transient assays, showing that IE0 is functional for very late gene activation and should be considered the 20th late gene expression factor (lef). In transient assays, IE0 showed that maximum very late gene expression is achieved at very low relative levels of protein. In contrast, IE1 requires higher levels of protein to obtain maximum very late gene expression. Furthermore, when the levels of IE0 become too high, very late gene expression rapidly declines. Interestingly, co-expression of IE0 and IE1 results in a mutually antagonistic affect on very late gene expression.